{"gene":"AKAP5","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":1996,"finding":"AKAP79 functions as a scaffold protein that simultaneously binds three signaling enzymes—PKA, calcineurin (PP2B), and PKC—at distinct binding sites, coordinating their localization in mammalian neurons.","method":"Deletion analysis and binding studies (co-immunoprecipitation, subcellular co-distribution by immunofluorescence)","journal":"Science","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal binding studies with deletion analysis, replicated across multiple subsequent labs","pmids":["8599116"],"is_preprint":false},{"year":1993,"finding":"AKAP79 (AKAP75) contains two distinct non-contiguous N-terminal domains (residues 27–48 and 77–91) required for intracellular membrane/cytoskeletal targeting, and a C-terminal RII-binding (tethering) domain mapped to residues 392–413 where hydrophobic residues are essential for high-affinity PKA regulatory subunit binding.","method":"Deletion and scanning mutagenesis with subcellular fractionation and RII binding assays in HEK293 cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — systematic mutagenesis with in vitro binding assays defining domain boundaries","pmids":["8509414"],"is_preprint":false},{"year":1997,"finding":"Ca²⁺/calmodulin binds AKAP79 with high affinity (KD ~28 nM) and competes with PKC for the same N-terminal region (residues 31–52), releasing inhibited PKC from the AKAP79 complex and increasing PKC activity at postsynaptic densities.","method":"Calmodulin binding assays, co-immunoprecipitation, PKC activity assays, immunofluorescence in hippocampal neurons","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro binding with KD measurement, functional PKC activity assay, cellular co-immunoprecipitation","pmids":["9202019"],"is_preprint":false},{"year":1998,"finding":"AKAP79 membrane targeting is mediated by three basic/hydrophobic N-terminal regions (A: residues 31–52; B: 76–101; C: 116–145) that bind acidic phospholipids including PtdIns(4,5)P2; this binding is regulated by phosphorylation and Ca²⁺/calmodulin, and PKC or calmodulin activation releases AKAP79 from membrane fractions.","method":"GFP-based fluorescence imaging of deletion mutants, membrane vesicle lipid-binding assays, subcellular fractionation, phosphorylation assays in HEK293 cells and cortical neurons","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (imaging, lipid binding, fractionation, phosphorylation) in a single study","pmids":["9545238"],"is_preprint":false},{"year":1998,"finding":"AKAP79 binds calcineurin A through residues 108–280 on AKAP79 and residues 30–98 and 311–336 on calcineurin A, independently of calcineurin B and at a site distinct from immunophilin-binding regions; overexpression of AKAP79 inhibits calcineurin-mediated NFAT dephosphorylation and activation in intact cells.","method":"Co-immunoprecipitation, truncation mapping, NFAT reporter assay in transfected cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — domain mapping with co-IP plus functional NFAT reporter assay in cells","pmids":["9765270"],"is_preprint":false},{"year":1999,"finding":"AKAP79 binds and inhibits the conserved catalytic core of PKC (multiple conventional, novel, and atypical isoforms) through a mechanism involving the pseudosubstrate displacement by residues 31–52 (specifically Arg39 and Arg40); lipid activators or kinase activation are not required for the association.","method":"In vitro binding and kinase activity assays, endoproteinase proteolysis, mutagenesis of AKAP79 and PKCβII, co-immunoprecipitation and immunofluorescence in hippocampal neurons","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with mutagenesis and multiple PKC isoforms tested","pmids":["10510312"],"is_preprint":false},{"year":2001,"finding":"AKAP79 regulates GRK2 membrane recruitment and β2AR phosphorylation by anchoring PKA, which directly phosphorylates GRK2 on Ser685, increasing Gβγ binding to GRK2 and enhancing GRK2 translocation to agonist-occupied receptor; S685A mutation or dominant-negative AKAP79 reduces GRK2-mediated receptor phosphorylation and internalization.","method":"Mutagenesis of GRK2 (S685A), dominant-negative AKAP79 expression, phosphorylation assays, receptor internalization assays in HEK293 cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — site-directed mutagenesis plus functional phosphorylation and internalization assays","pmids":["11278469"],"is_preprint":false},{"year":2001,"finding":"AKAP79 directly associates with the inwardly rectifying potassium channel Kir2.1 via both N- and C-terminal intracellular domains of the channel, and this association enhances the channel's response to elevated intracellular cAMP.","method":"Co-immunoprecipitation from intact cells, GST pulldown with Kir2.1 intracellular domains, electrophysiology in transfected cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — GST pulldown plus co-IP plus functional electrophysiology, single lab","pmids":["11287423"],"is_preprint":false},{"year":2002,"finding":"AKAP79 promotes basal PKA-mediated phosphorylation of GluR1 Ser845 and, through anchored PP2B and Ca²⁺ signaling, confers calcineurin-dependent downregulation of GluR1 receptor currents analogous to LTD; this requires PKA, Ser845, and PDZ-domain interaction between GluR1 and SAP97.","method":"Electrophysiology, co-immunoprecipitation, phosphorylation assays in HEK293 cells and hippocampal neurons","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal biochemistry plus functional electrophysiology with mutagenesis controls","pmids":["11943807"],"is_preprint":false},{"year":2002,"finding":"AKAP79 directly regulates cell-surface expression (trafficking) of L-type calcium channels (CaV1.2) via a polyproline sequence in the channel II–III cytoplasmic loop, independently of PKA activity.","method":"Extracellular epitope tagging of CaV1.2, immunoassays, whole-cell and single-channel electrophysiology in transfected cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional imaging plus electrophysiology, single lab","pmids":["12114507"],"is_preprint":false},{"year":2002,"finding":"The PP2B/calcineurin-binding site on AKAP79 is localized to residues 315–360; multiple determinants in this region bind directly to the calcineurin A subunit and inhibit phosphatase activity; peptides spanning residues 330–357 antagonize PP2B anchoring and attenuate PP2B-dependent downregulation of GluR1 currents.","method":"Cellular targeting assays with AKAP79 truncation/deletion mutants, in vitro binding and phosphatase activity assays, peptide antagonist electrophysiology in HEK293 cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — systematic mutagenesis with in vitro phosphatase assay plus functional electrophysiology","pmids":["12354762"],"is_preprint":false},{"year":2002,"finding":"FRET microscopy in living cells directly demonstrated that PKA-RII and calcineurin A subunits simultaneously bind AKAP79 at the plasma membrane cytoskeleton within ~5 nm of each other, forming a ternary kinase-scaffold-phosphatase complex; AKAP79 also regulates membrane localization of SAP97.","method":"FRET microscopy (immunofluorescence and live-cell FRET), subcellular localization in transfected cells","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — FRET in live cells providing direct distance measurement of ternary complex","pmids":["12507994"],"is_preprint":false},{"year":2003,"finding":"AKAP79 directly interacts with the C-terminal domain of IQGAP1, forming an IQGAP1/AKAP79 complex that co-purifies with PKA in β-cells, potentially linking cAMP/PKA signaling with Ca²⁺/CaM and GTPase pathways.","method":"cAMP affinity chromatography, co-immunoprecipitation, direct binding assay with IQGAP1 C-terminal domain","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — pulldown and co-IP, single lab, limited functional follow-up","pmids":["12938160"],"is_preprint":false},{"year":2005,"finding":"AKAP79 is constitutively associated with the β2-adrenergic receptor and anchors PKA to mediate PKA-dependent phosphorylation of β2AR and switching of β2AR signaling to ERK activation via Gi; β-arrestin-recruited PDE4D5 desensitizes this AKAP79/PKA-mediated process.","method":"siRNA knockdown of specific PDE4 isoforms and AKAP79, co-immunoprecipitation, β2AR phosphorylation assays, ERK activation assays in HEK293B2 cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — isoform-selective siRNA knockdown with multiple orthogonal functional readouts","pmids":["16030021"],"is_preprint":false},{"year":2006,"finding":"AKAP79-mediated targeting of PKA to the β1-adrenergic receptor promotes receptor recycling and functional resensitization; AKAP79, β1-AR, and PKA form a ternary complex at the β1-AR C-terminus; siRNA knockdown of AKAP79 prevents β1-AR recycling.","method":"siRNA knockdown, co-immunoprecipitation, FRET microscopy, PKA phosphorylation assays in HEK293 cells, SK-N-MC cells, and neonatal rat cortical neurons","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — siRNA rescue plus FRET plus phosphorylation assays, multiple cell types","pmids":["16940053"],"is_preprint":false},{"year":2006,"finding":"AKAP79 forms a complex with SAP97 at the PDZ-binding sequence (ESKV) at the β1-AR C-terminus, assembling a receptosome that targets PKA to β1-AR for Ser312 phosphorylation in the third intracellular loop, which is required for receptor recycling.","method":"Co-immunoprecipitation, domain deletion mutants, PKA phosphorylation assays, receptor trafficking assays in HEK293 cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with mutagenesis and functional trafficking readout, single lab","pmids":["17170109"],"is_preprint":false},{"year":2007,"finding":"AKAP79/150 directly interacts with CaV1.2 and co-targets PKA and calcineurin to L-type channels, conferring bidirectional regulation of L-type current; anchored calcineurin dominantly suppresses PKA enhancement of the channel; AKAP79/150-anchored calcineurin is required for NFATc4 activation by local Ca²⁺ influx through L-type channels.","method":"Co-immunoprecipitation, electrophysiology in HEK293 cells and hippocampal neurons, NFAT reporter assays, dominant-negative AKAP constructs","journal":"Neuron","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct binding plus electrophysiology plus transcription factor activation assay in two cell systems","pmids":["17640527"],"is_preprint":false},{"year":2008,"finding":"AKAP79/150 forms a complex with TRPV1, PKA, PKC, and calcineurin via a critical region in the TRPV1 C-terminus; disrupting this binding abrogates TRPV1 sensitization by bradykinin and PGE2, demonstrating that AKAP79/150 is a final common element for heat hyperalgesia.","method":"Co-immunoprecipitation, deletion mapping of TRPV1 C-terminus, TRPV1 electrophysiology in transfected cells and neurons, in vivo hyperalgesia assays","journal":"Neuron","confidence":"High","confidence_rationale":"Tier 2 / Strong — domain mapping, co-IP, functional electrophysiology, and in vivo behavioral assay","pmids":["18701070"],"is_preprint":false},{"year":2008,"finding":"AKAP79 selectively enhances PKC-mediated phosphorylation of GluR1 Ser831 by localizing PKC near the receptor via SAP97, shifting the PKC dose-dependence ~20-fold so that low PKC concentrations are as effective as much higher CaMKII concentrations.","method":"Biochemical phosphorylation assays, electrophysiology of GluR1 currents in transfected cells with AKAP79 constructs","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — biochemical and electrophysiological assays, single lab","pmids":["18305116"],"is_preprint":false},{"year":2010,"finding":"AKAP79 associates with multiple adenylyl cyclase (AC) isoforms (AC5, AC6, AC9) via their N-terminal regions and residues 77–108 of AKAP79; this interaction places AC5/6 in proximity to synaptic AMPA receptors; loss of AKAP150 in mice decreases AMPA receptor-associated AC activity.","method":"Co-immunoprecipitation, FRET (intensity- and lifetime-based) in living cells, AC activity assays in brain extracts from AKAP150 KO mice, domain deletion mapping","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — FRET in living cells plus biochemical assays plus mouse KO validation","pmids":["20231277"],"is_preprint":false},{"year":2010,"finding":"AKAP5 (AKAP79/150) organizes a caveolin-3-associated signaling complex in cardiac T-tubules comprising adenylyl cyclase 5/6, PKA, calcineurin, and a subpopulation of CaV1.2 channels; only this caveolin-3-associated CaV1.2 subpopulation is phosphorylated by PKA upon sympathetic stimulation; AKAP5 KO disrupts this complex, preventing normal calcium transients and PKA-dependent phosphorylation of ryanodine receptors and phospholamban.","method":"AKAP5 knockout mice, calcium imaging, electrophysiology, co-immunoprecipitation, PKA phosphorylation assays in cardiomyocytes","journal":"Circulation research","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO with multiple orthogonal biochemical and functional readouts","pmids":["20671242"],"is_preprint":false},{"year":2010,"finding":"AKAP79 alters the cellular pharmacology of anchored PKC, protecting it from certain ATP-competitive inhibitors; AKAP79-anchored PKC synchronizes muscarinic agonist-induced phosphorylation and KCNQ2 M-channel inhibition to optimize neuronal excitability.","method":"Dual fluorescent imaging/patch-clamp technique with CKAR kinase activity reporter, pharmacological profiling, electrophysiology in neurons","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — simultaneous live imaging and electrophysiology with pharmacological dissection, rigorous controls","pmids":["20188672"],"is_preprint":false},{"year":2010,"finding":"AKAP79/150 directly interacts with AC8, limiting AC8 sensitivity to intracellular Ca²⁺; this interaction was observed in HEK293 cells, pancreatic insulin-secreting cells, and hippocampal neurons.","method":"Co-immunoprecipitation, high-resolution live-cell imaging in multiple cell types endogenously expressing both proteins","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP in multiple physiologically relevant cell types, single lab","pmids":["20410303"],"is_preprint":false},{"year":2010,"finding":"AKAP5 knockout in mice delocalizes PKA from hippocampus and striatum (redistributing it to dendritic shafts via MAP2 binding); the D36 mutant lacking only the PKA binding domain produces more severe electrophysiological and behavioral deficits than complete KO, indicating that targeting calcineurin or other binding partners without the balancing PKA disrupts synaptic plasticity.","method":"Genetic KO and knockin (D36) mouse lines, immunofluorescence, co-immunoprecipitation, electrophysiology, behavioral tests","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with two distinct mutant lines plus multiple biochemical and functional readouts","pmids":["20428246"],"is_preprint":false},{"year":2010,"finding":"Ca²⁺/calmodulin disrupts AKAP79/150 interaction with KCNQ2–5 (but not KCNQ1) channels as shown by TIRF/FRET; this disruption prevents AKAP79-mediated sensitization of KCNQ2/3 channels to muscarinic inhibition, while PIP2 depletion does not affect AKAP79 membrane localization.","method":"TIRF/FRET microscopy, perforated patch-clamp electrophysiology, dominant-negative calmodulin, cotransfection in CHO cells","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — TIRF/FRET with electrophysiology and multiple molecular tools (WT vs DN CaM, PIP2 phosphatase)","pmids":["20147557"],"is_preprint":false},{"year":2011,"finding":"AKAP79 dimerizes through K328-K328 and K333-K333 cross-links; the reconstituted AKAP79-PP2B-RII-CaM complex has a molecular mass of ~466 kDa consisting of dimeric AKAP79 coordinating two RII homodimers, four PP2B heterodimers, and two calmodulin molecules; Ca²⁺/CaM binding activates anchored phosphatases by generating a second interface.","method":"Native mass spectrometry, chemical cross-linking, in vitro reconstitution of quaternary complex, quantitative biochemical analysis","journal":"Proceedings of the National Academy of Sciences","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with native MS and cross-linking providing stoichiometry and architecture","pmids":["21464287"],"is_preprint":false},{"year":2011,"finding":"AKAP79/150-anchored PKA controls Kv4.2 surface expression; the C-terminal domain of Kv4.2 interacts with an internal region of AKAP79/150 overlapping its MAGUK-binding domain; disrupting PKA anchoring decreases neuronal excitability while preventing calcineurin-mediated dephosphorylation increases excitability.","method":"Co-immunoprecipitation, pulldown, AKAP79/150 KO neurons, PKA anchoring disruption (Ht31 peptide), electrophysiology in hippocampal neurons","journal":"The Journal of neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — binding domain mapping plus functional electrophysiology, single lab","pmids":["21273417"],"is_preprint":false},{"year":2011,"finding":"AKAP79 is identified as a novel PP1 regulatory subunit; it directly binds PP1 catalytic subunit through a consensus FxxR/KxR/K motif in residues 1–44 and a second domain in residues 150–250; AKAP79 inhibits PP1 activity (IC50 ~811 nM) in a substrate-dependent manner but does not inhibit PP1 dephosphorylation of phospho-PSD-95.","method":"Co-immunoprecipitation from rat brain, pulldown with purified proteins, phosphatase activity assays, surface plasmon resonance, peptide mapping","journal":"Biochemistry","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — in vitro assays with purified proteins plus SPR, single lab","pmids":["21561082"],"is_preprint":false},{"year":2011,"finding":"Palmitoylation of AKAP79 at two N-terminal cysteines targets it to lipid rafts; mutation of these cysteines excludes AKAP79 from rafts, alters membrane diffusion behavior, and abolishes AKAP79-dependent regulation of SOCE-stimulated AC8 activity and PKA-dependent phosphorylation of raft proteins.","method":"Pharmacological palmitoylation inhibition, site-directed mutagenesis of Cys residues, lipid raft fractionation, FRAP analysis, AC8 activity assays in HEK293 cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — mutagenesis plus biochemical fractionation plus functional AC8 assay, multiple orthogonal methods","pmids":["21771783"],"is_preprint":false},{"year":2011,"finding":"AKAP79/150-mediated PKC anchoring is specifically required for muscarinic M1 (and angiotensin II) receptor suppression of M-type (KCNQ) K⁺ currents in SCG neurons, but not for bradykinin or purinergic suppression; FRET showed strong AKAP79 association with M1 and AT1 receptors and KCNQ2/3, but weak association with P2Y6 or B2 receptors.","method":"AKAP150 KO mice, transfection of ΔA-AKAP79 (no PKC binding), FRET under TIRF microscopy, perforated patch-clamp in SCG neurons","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO complemented by domain-specific mutant rescue plus FRET plus electrophysiology","pmids":["21562284"],"is_preprint":false},{"year":2012,"finding":"The IAIIIT anchoring motif in AKAP79 (residues forming a short linear motif) binds the same surface of calcineurin as the PxIxIT recognition peptide of NFAT but with higher affinity; increasing calcineurin-AKAP affinity paradoxically impairs NFAT activation by slowing calcineurin release and sequestering it as 'decoy' sites.","method":"Structural binding analysis, mutagenesis of AKAP79 anchoring sequence, NFAT activation assays, calcineurin recruitment measurements","journal":"Nature structural & molecular biology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — structural binding analysis plus mutagenesis plus functional NFAT assay revealing mechanistic balance","pmids":["22343722"],"is_preprint":false},{"year":2012,"finding":"AKAP79/150-anchored calcineurin and L-type Ca²⁺ channel activation drive KCNQ2/3 transcriptional upregulation via NFAT in hippocampal neurons; AKAP150 KO mice fail to upregulate KCNQ2/3 transcription after drug-induced seizures, indicating a negative feedback mechanism on neuronal excitability.","method":"Neuronal activity manipulation, NFAT reporter assays, AKAP150 KO mice, qRT-PCR of KCNQ2/3 mRNA in seizure model","journal":"Neuron","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO in physiological seizure model plus mechanistic reporter assays","pmids":["23259949"],"is_preprint":false},{"year":2012,"finding":"AKAP79 interacts with caldendrin (a neuronal Ca²⁺-binding protein) at a site overlapping the calmodulin-binding region; caldendrin competes with calmodulin for binding to AKAP79 with similar affinity (KD ~20 nM vs ~30 nM for CaM), but through an induced-fit mechanism with a slow rearrangement step and Ca²⁺-independent binding component.","method":"Pulldown experiments, surface plasmon resonance biosensor kinetic analysis","journal":"Journal of neurochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — SPR with kinetic modeling plus pulldown, single lab","pmids":["22693956"],"is_preprint":false},{"year":2012,"finding":"AKAP79 modulation of CaV1.2 membrane expression occurs through disruption of an autoinhibitory intramolecular interaction between the channel II–III linker and distal C-terminus; AKAP79 directly interacts with the distal CaV1.2 C-terminus, competing with its association to the II–III linker.","method":"Mutagenesis of polyproline domains, co-immunoprecipitation, channel trafficking assays in transfected cells","journal":"Channels","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mutagenesis plus co-IP plus functional channel expression assay, single lab","pmids":["22677788"],"is_preprint":false},{"year":2012,"finding":"PKA recruited to AC8 via AKAP79 phosphorylates AC8 at Ser112, providing a negative feedback mechanism that reduces the on-rate of cAMP production during Ca²⁺ oscillations.","method":"Site-directed mutagenesis of AC8 (S112A), FRET-based cAMP measurements during Ca²⁺ oscillations, co-immunoprecipitation","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1 / Strong — mutagenesis of substrate site plus live-cell FRET functional assay","pmids":["22976297"],"is_preprint":false},{"year":2013,"finding":"The AKAP79 region between amino acids 326–336 mediates binding to TRPV1; a TAT-conjugated peptide mimicking this domain inhibits TRPV1 sensitization in vitro and inflammatory hyperalgesia in vivo without affecting basal pain thresholds.","method":"FRET, co-immunoprecipitation, TRPV1 membrane trafficking assays, in vivo hyperalgesia assay with TAT-peptide","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — domain mapping with FRET/co-IP plus in vivo functional validation","pmids":["23699529"],"is_preprint":false},{"year":2013,"finding":"AKAP5 (AKAP79/150) anchors adenylyl cyclase (AC) to postsynaptic sites and this AC anchoring is required for β-adrenergic stimulation-induced phosphorylation of GluA1 Ser845 and for theta-frequency-induced LTP; AC anchoring (disrupted in AKAP5 KO) is more critical than PKA anchoring alone (disrupted in D36 mice) for this process.","method":"AKAP5 KO and D36 (PKA-binding-deleted) knock-in mice, phosphorylation assays, LTP electrophysiology in acute forebrain slices","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — two distinct genetic mouse models with biochemical and electrophysiological readouts","pmids":["23649627"],"is_preprint":false},{"year":2013,"finding":"AKAP79 recruits PKC to activate AC2 and generates localized cAMP upon Gq-coupled muscarinic receptor stimulation; PKA anchored to AKAP79 activates PDE4 to degrade this cAMP; calcineurin anchored to AKAP79 is not involved in this pathway.","method":"Live-cell cAMP imaging, siRNA knockdown of AKAP79 components, pharmacological dissection in HEK293 cells","journal":"The Biochemical journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — live-cell imaging plus siRNA dissection, single lab","pmids":["23889134"],"is_preprint":false},{"year":2014,"finding":"AKAP5 scaffold is required for PKA-GluA1 coupling during homeostatic plasticity; knockdown of AKAP5 blocks synaptic scaling up, which requires PKA-mediated phosphorylation of GluA1 S845; scaling down involves loss of PKA from synapses, and scaling up involves enhanced synaptic PKA activity regulated by AKAP5.","method":"AKAP5 RNAi knockdown, GluA1 S845 knockin mutant, phosphorylation assays, electrophysiology, surface AMPAR trafficking assays in neurons","journal":"Neuron","confidence":"High","confidence_rationale":"Tier 2 / Strong — knockin mutation plus RNAi plus phosphorylation and trafficking assays with functional electrophysiology","pmids":["25451194"],"is_preprint":false},{"year":2014,"finding":"AKAP5 controls calcineurin (CaN) and CaMKII activity in cardiac myocytes; loss of AKAP5 enhances CaN and CaMKII activities, interferes with β1-AR recycling through CaN binding to AKAP5, and leads to cardiac dilatation and dysfunction.","method":"AKAP5 KO mice, NFAT-luciferase reporter, echocardiography, biochemical assays, pharmacological rescue with carvedilol","journal":"Cardiovascular research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO with functional cardiac readouts and mechanistic reporter assay, single lab","pmids":["25225170"],"is_preprint":false},{"year":2015,"finding":"DHHC2 palmitoyl acyltransferase, resident in recycling endosomes, directly palmitoylates AKAP79/150 to regulate its targeting to recycling endosomes; DHHC2 knockdown disrupts recycling endosome exocytosis, dendritic spine enlargement, AKAP recruitment to spines, and LTP-stimulated AMPAR delivery; a palmitoylation-independent lipidated AKAP mutant rescues these deficits.","method":"RNAi knockdown of DHHC2, palmitoylation-independent AKAP rescue mutant, live imaging of spine exocytosis, electrophysiology in hippocampal neurons","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — RNAi with mechanistic rescue mutant plus multiple structural and functional plasticity readouts","pmids":["25589740"],"is_preprint":false},{"year":2015,"finding":"AKAP79 constitutively associates with plasma membrane STIM1 and mediates PKA phosphorylation of STIM1 Thr389, which is necessary for activation of store-independent ARC channels but actually inhibits STIM1's ability to activate store-operated CRAC channels.","method":"AKAP79 knockdown, STIM1 T389 mutagenesis, ARC and CRAC channel electrophysiology, PKA activity assays","journal":"The Journal of physiology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — site-directed mutagenesis of substrate residue plus functional electrophysiology distinguishing two channel types","pmids":["25504574"],"is_preprint":false},{"year":2017,"finding":"CaMKII mediates LTD-induced synaptic removal of AKAP79/150 by phosphorylating its N-terminal polybasic targeting domain (inhibiting F-actin association) and by promoting depalmitoylation at two N-terminal Cys residues; depalmitoylation (not phosphorylation per se) is required for AKAP79/150 spine removal and LTD-induced spine shrinkage; autonomous CaMKII activity preferentially phosphorylates AKAP79/150 compared with Ca²⁺/CaM-stimulated CaMKII.","method":"CaMKII inhibitors and constitutively active CaMKII, mutagenesis of palmitoylation sites and phosphorylation sites, live imaging of spine morphology, AKAP trafficking in hippocampal neurons during LTD","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple mutagenesis approaches plus pharmacological dissection plus live imaging with functional LTD readout","pmids":["29196604"],"is_preprint":false},{"year":2017,"finding":"Intrinsic disorder in AKAP79 allows conformational flexibility in PP2B engagement; the sole PP2B-anchoring determinant is a short linear motif (residues 337–343); Ca²⁺/calmodulin activation condenses diverse conformational variants into a uniform 178 Å population and engages a Leu-Lys-Ile-Pro sequence (residues 125–128) that occupies a PP2B binding pocket shared with cyclosporin, fine-tuning anchored phosphatase drug sensitivity.","method":"Negative-stain electron microscopy structural analysis, live-cell fluorescent activity sensor imaging, mutagenesis","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1 / Strong — EM structural analysis plus live-cell functional imaging plus mutagenesis in single study","pmids":["28967377"],"is_preprint":false},{"year":2019,"finding":"AKAP79 directly interacts with NFAT via its C-terminal leucine-zipper (LZ) domain; disrupting this LZ interaction abolishes depolarization-stimulated NFAT signaling in hippocampal neurons while preserving the AKAP-LTCC interaction and LTCC function; AKAP79 thus recruits NFAT to the LTCC signaling complex to promote its activation by anchored calcineurin.","method":"RNAi knockdown of AKAP150 with human AKAP79 LZ-mutant replacement, FRET, Ca²⁺ imaging, electrophysiology, NFAT reporter, FRAP and FCS in hippocampal neurons","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — domain-specific mutant rescue plus FRET plus multiple functional assays in neurons","pmids":["31091162"],"is_preprint":false},{"year":2019,"finding":"AKAP79-anchored PKC (not other AKAP79-signaling components) drives the appearance of Ca²⁺-permeable (GluA2-lacking) AMPARs primarily through GluA1 Ser831 phosphorylation; this generates CP-AMPARs under conditions where CI-AMPARs normally predominate.","method":"Electrophysiology (current-voltage relationships), GluA1 phosphosite mutagenesis, AKAP79 domain mutants in transfected cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mutagenesis plus electrophysiology, single lab","pmids":["30737285"],"is_preprint":false},{"year":2020,"finding":"AKAP5 organizes a nanocomplex of P2Y11/P2Y11-like receptors, AC5, PKA, and CaV1.2 at the plasma membrane of arterial myocytes; disruption of AKAP5 function blocks glucose- and P2Y11 agonist-induced cAMP synthesis, CaV1.2 potentiation, vasoconstriction, and decreased blood flow.","method":"AKAP5 null mice and arterial myocytes, proximity ligation assay for nanocomplexes, cAMP assays, electrophysiology, vasoconstriction assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO plus proximity ligation plus multiple functional vascular readouts","pmids":["33082339"],"is_preprint":false},{"year":2020,"finding":"STIM2 recruits Orai1/STIM1 to ER-PM junctions in response to ER-Ca²⁺ depletion, promoting assembly with AKAP79 to couple Orai1 channel function to NFAT1 activation; STIM2 knockdown attenuates NFAT1 activation and Orai1-AKAP79 assembly without substantially reducing Orai1/STIM1 clustering or global Ca²⁺ increases.","method":"STIM2 knockdown, STIM1ΔK mutant, co-immunoprecipitation, Ca²⁺ imaging, NFAT1 nuclear translocation assays","journal":"Proceedings of the National Academy of Sciences","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockdown plus multiple mutant constructs plus biochemical and functional Ca²⁺/NFAT assays","pmids":["32601188"],"is_preprint":false},{"year":2021,"finding":"AKAP79 enables calcineurin to dephosphorylate PKA type II regulatory subunits at a rate ~10-fold higher than without scaffolding; this allows calcineurin to suppress PKA activity by increasing catalytic subunit capture rate without altering cAMP levels, and this mechanism contributes to hippocampal LTD.","method":"In vitro reconstitution phosphatase assays, fluorescent PKA activity reporter (AKAR), kinetic modeling, hippocampal neuron LTD electrophysiology","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution plus live-cell PKA reporter plus kinetic modeling plus LTD electrophysiology","pmids":["34612814"],"is_preprint":false},{"year":2021,"finding":"The N-terminus of Orai1 (but not Orai2, Orai3, or a shorter Orai1 isoform) directly interacts with AKAP79; NMR structural analysis reveals the AKAP-binding domain has a compact shape with proline-driven turns; this interaction is essential for NFAT1 activation by local Ca²⁺ entry and cytokine production.","method":"NMR structural analysis of AKAP79-binding domain, co-immunoprecipitation, domain deletion/truncation of Orai isoforms, NFAT1 reporter assays, cytokine production assays","journal":"Proceedings of the National Academy of Sciences","confidence":"High","confidence_rationale":"Tier 1 / Strong — NMR structure plus biochemical binding mapping plus multiple functional assays","pmids":["33941685"],"is_preprint":false},{"year":2021,"finding":"AKAP79 coordinates PKA and PDE4 in a cAMP signaling nexus adjacent to Orai1; both PKA and PDE4 associate with AKAP79 and relocalize close to Orai1 after stimulation; in HEK293 cells, which lack functional Ca²⁺-activated adenylyl cyclases including AC8, Ca²⁺ entry through Orai1 does not increase cAMP levels despite AKAP79-Orai1 association.","method":"FRET-based cAMP sensors (AKAP79-CUTie), mass spectrometry, PCR, bulk cAMP and PKA activity measurements in HEK293 cells","journal":"Function","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — targeted FRET reporter plus mass spectrometry with multiple negative controls, single lab","pmids":["34458850"],"is_preprint":false},{"year":2021,"finding":"AKAP79/150 coordinates leptin-induced PKA signaling at the cell membrane in pancreatic β-cells; leptin increases PKA activity at the membrane via NMDAR-CaMKKβ-AMPK signaling in an AKAP79/150-dependent manner; disrupting PP2B anchoring to AKAP79/150 elevates basal PKA signaling and increases surface KATP channels even without leptin.","method":"FRET-based PKA activity reporters, AKAP79/150 genetic knockdown and rescue, dominant-negative PP2B anchoring disruption, KATP channel trafficking assays in MIN6 β-cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — FRET reporter plus genetic knockdown rescue plus channel trafficking assay, single lab","pmids":["33617875"],"is_preprint":false},{"year":2010,"finding":"A pre-assembled RXFP1-AKAP79-AC2-β-arrestin 2-PDE4D3 signalosome mediates sub-picomolar relaxin signaling; AC2 is functionally coupled to RXFP1 through AKAP79 binding to helix 8 of the receptor; PKA-activated PDE4D3 scaffolded via β-arrestin 2 tonically opposes AC2 activity.","method":"cAMP biosensors in single cells, co-immunoprecipitation, domain mapping (helix 8 of RXFP1)","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — live-cell cAMP imaging plus biochemical co-IP with domain identification, single lab","pmids":["20664520"],"is_preprint":false},{"year":2022,"finding":"The AKAP79-Orai1 interaction is considerably more transient than STIM1-Orai1; free AKAP79 (with calcineurin and NFAT1) can rapidly replace AKAP79 devoid of NFAT1 on Orai1 during continuous Ca²⁺ entry; Ca²⁺ nanodomains near Orai1 activate nearly the entire cytosolic NFAT1 pool, and recycling of inactive NFAT1 from cytoplasm to AKAP79 sustains excitation-transcription coupling.","method":"FRAP, FCS, co-immunoprecipitation, NFAT1 translocation assays, mathematical kinetic modeling","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — FRAP/FCS kinetic measurements plus mathematical model, single lab","pmids":["36317924"],"is_preprint":false}],"current_model":"AKAP79/150 (AKAP5) is a multivalent postsynaptic scaffold protein that simultaneously anchors PKA, calcineurin (PP2B), PKC, PP1, and adenylyl cyclase isoforms (AC5/6/8/9) at the plasma membrane via distinct binding domains; it targets these enzymes to ion channels (CaV1.2, TRPV1, KCNQ2-5, Kir2.1, Kv4.2, KATP, HERG), GPCRs (β1-AR, β2-AR, RXFP1, GPR30), and AMPA receptors through direct protein-protein interactions or scaffold intermediaries (SAP97, caveolin-3), thereby organizing compartmentalized phosphorylation/dephosphorylation cycles that bidirectionally regulate synaptic plasticity, neuronal excitability, channel trafficking, NFAT-dependent transcription, and cardiac β-adrenergic signaling; its membrane targeting is determined by lipid (PtdIns(4,5)P2) binding through polybasic N-terminal regions and palmitoylation at two N-terminal cysteines (mediated by DHHC2), while LTD-stimulus-induced CaMKII phosphorylation and depalmitoylation drive its synaptic removal."},"narrative":{"mechanistic_narrative":"AKAP5 (AKAP79/150) is a multivalent membrane-targeted scaffold that simultaneously anchors the kinase PKA, the phosphatase calcineurin (PP2B), and PKC at distinct binding sites to coordinate compartmentalized phosphorylation/dephosphorylation cycles in neurons and other tissues [PMID:8599116]. Systematic mutagenesis defined its modular architecture: a C-terminal RII (PKA)-binding helix [PMID:8509414], an internal calcineurin-anchoring site centered on a short IAIIIT/LxxIP linear motif [PMID:12354762, PMID:22343722, PMID:28967377], and an N-terminal region that both binds and inhibits the PKC catalytic core via a pseudosubstrate-like Arg39/Arg40 mechanism [PMID:10510312]. The same N-terminal polybasic/hydrophobic regions target the scaffold to the membrane through PtdIns(4,5)P2 and acidic phospholipid binding [PMID:9545238] and through DHHC2-mediated palmitoylation of two N-terminal cysteines that direct it to lipid rafts and recycling endosomes [PMID:21771783, PMID:25589740]; Ca2+/calmodulin and PKC activation release AKAP5 from membranes and from channel partners, and calmodulin competes directly with PKC for the N-terminal site [PMID:9202019, PMID:9545238, PMID:20147557]. Native mass spectrometry and EM established that AKAP5 dimerizes to assemble a ~466 kDa quaternary complex in which Ca2+/CaM binding generates a second interface that activates anchored phosphatase, and that intrinsic disorder permits conformational flexibility in PP2B engagement [PMID:21464287, PMID:28967377]. Through these modules AKAP5 docks PKA, calcineurin, PKC, adenylyl cyclases (AC2/5/6/8/9) and PP1 onto ion channels and receptors—CaV1.2, TRPV1, KCNQ2-5, Kir2.1, Kv4.2, KATP, Orai1/STIM, and β1/β2-adrenergic, muscarinic, P2Y and RXFP1 GPCRs—to bidirectionally control channel activity, trafficking and receptor recycling [PMID:17640527, PMID:18701070, PMID:20147557, PMID:21273417, PMID:33082339, PMID:33941685, PMID:20664520]. In neurons this organization governs AMPA receptor phosphorylation and trafficking underlying LTP, LTD and homeostatic synaptic scaling [PMID:11943807, PMID:23649627, PMID:25451194, PMID:34612814], and couples local Ca2+ entry through L-type channels and Orai1 to calcineurin-dependent NFAT transcription, including feedback upregulation of KCNQ channels [PMID:17640527, PMID:23259949, PMID:31091162, PMID:33941685]. In cardiac and arterial myocytes AKAP5 nucleates caveolin-3- and AC5-associated nanocomplexes that confer β-adrenergic and purinergic control of CaV1.2, calcium handling, and vasoconstriction, with knockout causing cardiac dilatation and dysfunction [PMID:20671242, PMID:25225170, PMID:33082339]. AKAP5 also functions as a PKA-dependent regulatory node for GRK2 recruitment and β2AR desensitization [PMID:11278469] and acts directly as a PP1 inhibitory subunit [PMID:21561082].","teleology":[{"year":1993,"claim":"Established the modular domain organization that makes AKAP5 a targeting scaffold, separating membrane/cytoskeletal anchoring from PKA tethering.","evidence":"Deletion and scanning mutagenesis with subcellular fractionation and RII binding assays in HEK293 cells","pmids":["8509414"],"confidence":"High","gaps":["Did not address how the same protein engages calcineurin or PKC","Membrane targets of the N-terminal domains not yet identified"]},{"year":1996,"claim":"Defined the central concept that AKAP5 is a single scaffold binding three signaling enzymes (PKA, calcineurin, PKC) at distinct sites, enabling coordinated localization.","evidence":"Deletion analysis and binding studies with co-IP and immunofluorescence co-distribution in neurons","pmids":["8599116"],"confidence":"High","gaps":["Whether the three enzymes occupy one complex simultaneously not directly shown","No functional readout of coordinated signaling"]},{"year":1997,"claim":"Showed Ca2+/calmodulin acts as a regulatory switch on the scaffold by competing with PKC for the N-terminal site and releasing active PKC.","evidence":"Calmodulin binding assays with KD measurement, co-IP, PKC activity assays in hippocampal neurons","pmids":["9202019"],"confidence":"High","gaps":["Physiological trigger of CaM competition in vivo not established"]},{"year":1998,"claim":"Resolved how AKAP5 reaches the membrane and how PKC inhibition works, linking lipid binding and pseudosubstrate displacement to regulated targeting.","evidence":"GFP imaging of mutants, lipid-binding/fractionation assays, in vitro PKC binding and kinase assays with mutagenesis (combined across studies)","pmids":["9545238","9765270","10510312"],"confidence":"High","gaps":["Relative in vivo contribution of lipid binding vs palmitoylation unresolved at this stage","Calcineurin inhibition mechanism only partially mapped"]},{"year":2002,"claim":"Demonstrated that the scaffold assembles a true ternary kinase-scaffold-phosphatase complex and uses it to bidirectionally regulate AMPA receptors and L-type channels.","evidence":"Live-cell FRET (~5 nm), electrophysiology, NFAT reporter and phosphorylation assays in HEK293 and hippocampal neurons (multiple studies)","pmids":["12507994","11943807","12114507","12354762"],"confidence":"High","gaps":["Stoichiometry and oligomeric state of the complex not yet defined","Trafficking effects on CaV1.2 PKA-independent — mechanism unclear at this point"]},{"year":2007,"claim":"Connected the scaffold to excitation-transcription coupling, showing anchored calcineurin reads local L-type channel Ca2+ to activate NFAT.","evidence":"Co-IP, electrophysiology and NFAT reporter assays with dominant-negative AKAP in HEK293 and neurons","pmids":["17640527"],"confidence":"High","gaps":["How NFAT itself is recruited to the complex not yet defined"]},{"year":2008,"claim":"Identified AKAP5 as the convergent platform for inflammatory TRPV1 sensitization, extending its role to nociception.","evidence":"Co-IP, TRPV1 C-terminus deletion mapping, electrophysiology and in vivo hyperalgesia assays","pmids":["18701070"],"confidence":"High","gaps":["Relative contribution of each anchored enzyme to sensitization not dissected here"]},{"year":2010,"claim":"Broadened the scaffold's enzyme repertoire to adenylyl cyclases and extended its role to cardiac and GPCR signalosomes, establishing tissue-wide compartmentalized cAMP control.","evidence":"Co-IP, FRET, AC activity assays in AKAP150 KO brain, cardiac KO mice, and cAMP biosensors (multiple studies)","pmids":["20231277","20671242","20410303","20188672","20428246","20664520"],"confidence":"High","gaps":["How AC isoform selectivity is achieved across tissues not fully mapped","D36 vs full-KO phenotype divergence mechanism partly unexplained"]},{"year":2011,"claim":"Defined the quaternary architecture and added PP1 regulation and palmitoylation-dependent raft targeting, explaining how the scaffold physically integrates multiple enzymes and is membrane-positioned.","evidence":"Native MS and cross-linking of reconstituted complex, SPR/phosphatase assays, palmitoylation mutagenesis with raft fractionation and AC8 assays (multiple studies)","pmids":["21464287","21561082","21771783","21273417","21562284"],"confidence":"High","gaps":["In vivo relevance of PP1 inhibition not established","Which channel/receptor complexes depend on raft localization not fully cataloged"]},{"year":2012,"claim":"Revealed a paradoxical affinity-tuning principle: the calcineurin-anchoring motif competes with NFAT's docking peptide, so AKAP5 affinity sets NFAT output, and PKA feedback phosphorylates AC8 to shape cAMP dynamics.","evidence":"Structural binding analysis with mutagenesis, NFAT/calcineurin recruitment assays, AC8 S112A mutagenesis with FRET cAMP imaging (multiple studies)","pmids":["22343722","22976297","23259949","22677788","22693956"],"confidence":"High","gaps":["Whether decoy-site tuning operates at endogenous expression levels unresolved"]},{"year":2013,"claim":"Established AC anchoring (not just PKA anchoring) as the critical node for β-adrenergic GluA1 phosphorylation and LTP, and validated the TRPV1-binding motif as an analgesic target in vivo.","evidence":"AKAP5 KO and D36 knock-in mice with LTP electrophysiology; TAT-peptide hyperalgesia assays (multiple studies)","pmids":["23649627","23699529","23889134"],"confidence":"High","gaps":["Mechanism by which AC anchoring outweighs PKA anchoring not fully explained"]},{"year":2014,"claim":"Showed AKAP5 governs homeostatic synaptic scaling and cardiac CaN/CaMKII balance, broadening its plasticity and disease roles.","evidence":"AKAP5 RNAi and GluA1 S845 knockin, trafficking/electrophysiology; AKAP5 KO mice with echocardiography (multiple studies)","pmids":["25451194","25225170"],"confidence":"High","gaps":["Causal chain from AKAP5 loss to cardiac dilatation only partly resolved"]},{"year":2015,"claim":"Identified DHHC2 as the enzyme controlling AKAP5 palmitoylation/endosomal targeting and showed scaffold coupling to STIM1/store-operated channels, linking lipid modification to plasticity and Ca2+ entry.","evidence":"DHHC2 RNAi with lipidated rescue mutant, spine imaging, electrophysiology; STIM1 T389 mutagenesis with ARC/CRAC recordings (multiple studies)","pmids":["25589740","25504574"],"confidence":"High","gaps":["Regulation of DHHC2 activity itself unknown","How STIM1 phosphorylation discriminates ARC vs CRAC at structural level not defined"]},{"year":2017,"claim":"Defined the molecular switch for activity-dependent scaffold removal, showing CaMKII-driven depalmitoylation (not phosphorylation alone) drives synaptic removal during LTD, and that intrinsic disorder tunes anchored phosphatase drug sensitivity.","evidence":"CaMKII pharmacology and site mutagenesis with live spine imaging; negative-stain EM with live-cell sensors (multiple studies)","pmids":["29196604","28967377"],"confidence":"High","gaps":["Identity of the depalmitoylating enzyme not established","How conformational ensembles map to specific functional states unresolved"]},{"year":2019,"claim":"Pinpointed the C-terminal leucine-zipper as the direct NFAT-recruitment determinant and assigned PKC-dependent GluA1 Ser831 phosphorylation to generation of Ca2+-permeable AMPARs.","evidence":"AKAP150 RNAi with LZ-mutant rescue, FRET/NFAT/Ca2+ assays; GluA1 phosphosite mutagenesis with I-V electrophysiology (multiple studies)","pmids":["31091162","30737285"],"confidence":"High","gaps":["Structural basis of LZ-NFAT contact not solved"]},{"year":2021,"claim":"Quantified how scaffolding accelerates calcineurin-mediated suppression of PKA and dissected the Orai1-AKAP5-NFAT cytokine pathway, giving a kinetic and structural account of excitation-transcription coupling and PKA antagonism.","evidence":"In vitro reconstitution phosphatase assays with PKA reporters and LTD electrophysiology; NMR of the Orai1 N-terminus; cAMP nexus mapping; β-cell leptin FRET reporters (multiple studies)","pmids":["34612814","33941685","34458850","33617875","33082339"],"confidence":"High","gaps":["Whether the ~10-fold calcineurin acceleration occurs at endogenous stoichiometry in vivo not confirmed","AC requirement for Orai1-linked cAMP appears cell-type-dependent"]},{"year":2022,"claim":"Established dynamic exchange kinetics of AKAP5 on Orai1, explaining how transient scaffold occupancy and NFAT recycling sustain transcription during continuous Ca2+ entry.","evidence":"FRAP, FCS, co-IP and NFAT1 translocation with mathematical kinetic modeling","pmids":["36317924"],"confidence":"Medium","gaps":["Single-lab kinetic model awaits independent confirmation","Molecular driver of the rapid AKAP5 exchange not identified"]},{"year":null,"claim":"It remains unresolved how the many enzyme and channel modules are dynamically prioritized at a single AKAP5 scaffold within a cell, and which depalmitoylase and upstream signals control the activity-dependent assembly/disassembly cycle in vivo.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No full-length high-resolution structure of an assembled signalosome on a native channel","Depalmitoylating enzyme unidentified","Selectivity rules governing which partner complex forms in a given compartment are unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,11,25,44]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[5,10,27,30,48]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[3,28]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[3,42]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3,11,20,46]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[40]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[1,11,42]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[23,53]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,6,13,14,37,52]},{"term_id":"R-HSA-112316","term_label":"Neuronal System","supporting_discovery_ids":[8,17,36,38,42]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[16,31,44,49]},{"term_id":"R-HSA-397014","term_label":"Muscle contraction","supporting_discovery_ids":[20,39,46]}],"complexes":["AKAP79-PKA(RII)-calcineurin(PP2B)-CaM quaternary complex","caveolin-3/AC5-6/PKA/calcineurin/CaV1.2 cardiac T-tubule complex","RXFP1-AKAP79-AC2-β-arrestin2-PDE4D3 signalosome","Orai1-AKAP79-calcineurin-NFAT1 complex"],"partners":["PRKAR2 (PKA RII)","PPP3CA (CALCINEURIN A)","PRKCB (PKC)","CALM1 (CALMODULIN)","ADCY8 (AC8)","DLG1 (SAP97)","CACNA1C (CAV1.2)","ORAI1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P24588","full_name":"A-kinase anchor protein 5","aliases":["A-kinase anchor protein 79 kDa","AKAP 79","H21","cAMP-dependent protein kinase regulatory subunit II high affinity-binding protein"],"length_aa":427,"mass_kda":47.1,"function":"Multivalent scaffold protein that anchors the cAMP-dependent protein kinase/PKA to cytoskeletal and/or organelle-associated proteins, targeting the signal carried by cAMP to specific intracellular effectors (PubMed:1512224). Association with the beta2-adrenergic receptor (beta2-AR) not only regulates beta2-AR signaling pathway, but also the activation by PKA by switching off the beta2-AR signaling cascade. Plays a role in long term synaptic potentiation by regulating protein trafficking from the dendritic recycling endosomes to the plasma membrane and controlling both structural and functional plasticity at excitatory synapses (PubMed:25589740). In hippocampal pyramidal neurons, recruits KCNK2/TREK-1 channel at postsynaptic dense bodies microdomains and converts it to a leak channel no longer sensitive to stimulation by arachidonic acid, acidic pH or mechanical stress, nor inhibited by Gq-coupled receptors but still under the negative control of Gs-coupled receptors (By similarity). Associates with ORAI1 pore-forming subunit of CRAC channels in Ca(2+) signaling microdomains where it recruits NFATC2/NFAT1 and couples store-operated Ca(2+) influx to calmodulin and calcineurin signaling and activation of NFAT-dependent transcriptional responses (PubMed:33941685)","subcellular_location":"Postsynaptic recycling endosome membrane; Cell projection, dendrite; Postsynaptic cell membrane","url":"https://www.uniprot.org/uniprotkb/P24588/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/AKAP5","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"PRKACA","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/AKAP5","total_profiled":1310},"omim":[{"mim_id":"616427","title":"A-KINASE ANCHOR INHIBITOR 1; AKAIN1","url":"https://www.omim.org/entry/616427"},{"mim_id":"604688","title":"A-KINASE ANCHOR PROTEIN 5; AKAP5","url":"https://www.omim.org/entry/604688"},{"mim_id":"603870","title":"CORE-BINDING FACTOR, ALPHA SUBUNIT 2, TRANSLOCATED TO, 3; CBFA2T3","url":"https://www.omim.org/entry/603870"},{"mim_id":"114105","title":"PROTEIN PHOSPHATASE 3, CATALYTIC SUBUNIT, ALPHA ISOFORM; PPP3CA","url":"https://www.omim.org/entry/114105"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Golgi apparatus","reliability":"Approved"},{"location":"Plasma membrane","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"brain","ntpm":17.6}],"url":"https://www.proteinatlas.org/search/AKAP5"},"hgnc":{"alias_symbol":["AKAP75","AKAP79"],"prev_symbol":[]},"alphafold":{"accession":"P24588","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P24588","model_url":"https://alphafold.ebi.ac.uk/files/AF-P24588-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P24588-F1-predicted_aligned_error_v6.png","plddt_mean":54.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=AKAP5","jax_strain_url":"https://www.jax.org/strain/search?query=AKAP5"},"sequence":{"accession":"P24588","fasta_url":"https://rest.uniprot.org/uniprotkb/P24588.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P24588/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P24588"}},"corpus_meta":[{"pmid":"8599116","id":"PMC_8599116","title":"Coordination of three signaling enzymes by AKAP79, a mammalian scaffold protein.","date":"1996","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/8599116","citation_count":479,"is_preprint":false},{"pmid":"17640527","id":"PMC_17640527","title":"AKAP79/150 anchoring of calcineurin controls neuronal L-type Ca2+ channel activity and nuclear signaling.","date":"2007","source":"Neuron","url":"https://pubmed.ncbi.nlm.nih.gov/17640527","citation_count":285,"is_preprint":false},{"pmid":"18701070","id":"PMC_18701070","title":"Proinflammatory mediators modulate the heat-activated ion channel TRPV1 via the scaffolding protein AKAP79/150.","date":"2008","source":"Neuron","url":"https://pubmed.ncbi.nlm.nih.gov/18701070","citation_count":218,"is_preprint":false},{"pmid":"9545238","id":"PMC_9545238","title":"Membrane-targeting sequences on AKAP79 bind phosphatidylinositol-4, 5-bisphosphate.","date":"1998","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/9545238","citation_count":204,"is_preprint":false},{"pmid":"11943807","id":"PMC_11943807","title":"Regulation of GluR1 by the A-kinase anchoring protein 79 (AKAP79) signaling complex shares properties with long-term depression.","date":"2002","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/11943807","citation_count":183,"is_preprint":false},{"pmid":"16030021","id":"PMC_16030021","title":"RNA silencing identifies PDE4D5 as the functionally relevant cAMP phosphodiesterase interacting with beta arrestin to control the protein kinase A/AKAP79-mediated switching of the beta2-adrenergic receptor to activation of ERK in HEK293B2 cells.","date":"2005","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16030021","citation_count":170,"is_preprint":false},{"pmid":"20671242","id":"PMC_20671242","title":"Sympathetic stimulation of adult cardiomyocytes requires association of AKAP5 with a subpopulation of L-type calcium channels.","date":"2010","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/20671242","citation_count":141,"is_preprint":false},{"pmid":"11278469","id":"PMC_11278469","title":"Regulation of membrane targeting of the G protein-coupled receptor kinase 2 by protein kinase A and its anchoring protein AKAP79.","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11278469","citation_count":135,"is_preprint":false},{"pmid":"25451194","id":"PMC_25451194","title":"PKA-GluA1 coupling via AKAP5 controls AMPA receptor phosphorylation and cell-surface targeting during bidirectional homeostatic plasticity.","date":"2014","source":"Neuron","url":"https://pubmed.ncbi.nlm.nih.gov/25451194","citation_count":132,"is_preprint":false},{"pmid":"22343722","id":"PMC_22343722","title":"Balanced interactions of calcineurin with AKAP79 regulate Ca2+-calcineurin-NFAT signaling.","date":"2012","source":"Nature structural & molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/22343722","citation_count":130,"is_preprint":false},{"pmid":"12354762","id":"PMC_12354762","title":"Mapping the protein phosphatase-2B anchoring site on AKAP79. Binding and inhibition of phosphatase activity are mediated by residues 315-360.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12354762","citation_count":124,"is_preprint":false},{"pmid":"10878251","id":"PMC_10878251","title":"AKAP79 and the evolution of the AKAP model.","date":"2000","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/10878251","citation_count":112,"is_preprint":false},{"pmid":"12114507","id":"PMC_12114507","title":"Trafficking of L-type calcium channels mediated by the postsynaptic scaffolding protein AKAP79.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12114507","citation_count":112,"is_preprint":false},{"pmid":"12507994","id":"PMC_12507994","title":"Imaging kinase--AKAP79--phosphatase scaffold complexes at the plasma membrane in living cells using FRET microscopy.","date":"2002","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/12507994","citation_count":109,"is_preprint":false},{"pmid":"9202019","id":"PMC_9202019","title":"Regulation of the AKAP79-protein kinase C interaction by Ca2+/Calmodulin.","date":"1997","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9202019","citation_count":99,"is_preprint":false},{"pmid":"20188672","id":"PMC_20188672","title":"Interaction with AKAP79 modifies the cellular pharmacology of PKC.","date":"2010","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/20188672","citation_count":96,"is_preprint":false},{"pmid":"20231277","id":"PMC_20231277","title":"AKAP79 interacts with multiple adenylyl cyclase (AC) isoforms and scaffolds AC5 and -6 to alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) receptors.","date":"2010","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20231277","citation_count":95,"is_preprint":false},{"pmid":"8509414","id":"PMC_8509414","title":"Characterization of distinct tethering and intracellular targeting domains in AKAP75, a protein that links cAMP-dependent protein kinase II beta to the cytoskeleton.","date":"1993","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/8509414","citation_count":79,"is_preprint":false},{"pmid":"21464287","id":"PMC_21464287","title":"Architecture and dynamics of an A-kinase anchoring protein 79 (AKAP79) signaling complex.","date":"2011","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/21464287","citation_count":78,"is_preprint":false},{"pmid":"10510312","id":"PMC_10510312","title":"Mechanism of A-kinase-anchoring protein 79 (AKAP79) and protein kinase C interaction.","date":"1999","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/10510312","citation_count":75,"is_preprint":false},{"pmid":"21771783","id":"PMC_21771783","title":"Palmitoylation targets AKAP79 protein to lipid rafts and promotes its regulation of calcium-sensitive adenylyl cyclase type 8.","date":"2011","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21771783","citation_count":74,"is_preprint":false},{"pmid":"20428246","id":"PMC_20428246","title":"Mutations in AKAP5 disrupt dendritic signaling complexes and lead to electrophysiological and behavioral phenotypes in mice.","date":"2010","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/20428246","citation_count":73,"is_preprint":false},{"pmid":"9765270","id":"PMC_9765270","title":"AKAP79 inhibits calcineurin through a site distinct from the immunophilin-binding region.","date":"1998","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9765270","citation_count":72,"is_preprint":false},{"pmid":"23259949","id":"PMC_23259949","title":"Activity-dependent transcriptional regulation of M-Type (Kv7) K(+) channels by AKAP79/150-mediated NFAT actions.","date":"2012","source":"Neuron","url":"https://pubmed.ncbi.nlm.nih.gov/23259949","citation_count":72,"is_preprint":false},{"pmid":"16940053","id":"PMC_16940053","title":"AKAP79-mediated targeting of the cyclic AMP-dependent protein kinase to the beta1-adrenergic receptor promotes recycling and functional resensitization of the receptor.","date":"2006","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16940053","citation_count":71,"is_preprint":false},{"pmid":"20664520","id":"PMC_20664520","title":"Sub-picomolar relaxin signalling by a pre-assembled RXFP1, AKAP79, AC2, beta-arrestin 2, PDE4D3 complex.","date":"2010","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/20664520","citation_count":67,"is_preprint":false},{"pmid":"20410303","id":"PMC_20410303","title":"AKAP79/150 interacts with AC8 and regulates Ca2+-dependent cAMP synthesis in pancreatic and neuronal systems.","date":"2010","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20410303","citation_count":66,"is_preprint":false},{"pmid":"17170109","id":"PMC_17170109","title":"Assembly of an SAP97-AKAP79-cAMP-dependent protein kinase scaffold at the type 1 PSD-95/DLG/ZO1 motif of the human beta(1)-adrenergic receptor generates a receptosome involved in receptor recycling and networking.","date":"2006","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17170109","citation_count":65,"is_preprint":false},{"pmid":"25589740","id":"PMC_25589740","title":"The palmitoyl acyltransferase DHHC2 regulates recycling endosome exocytosis and synaptic potentiation through palmitoylation of AKAP79/150.","date":"2015","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/25589740","citation_count":61,"is_preprint":false},{"pmid":"23649627","id":"PMC_23649627","title":"Adenylyl cyclase anchoring by a kinase anchor protein AKAP5 (AKAP79/150) is important for postsynaptic β-adrenergic signaling.","date":"2013","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/23649627","citation_count":58,"is_preprint":false},{"pmid":"11287423","id":"PMC_11287423","title":"Targeting of an A kinase-anchoring protein, AKAP79, to an inwardly rectifying potassium channel, Kir2.1.","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11287423","citation_count":56,"is_preprint":false},{"pmid":"24962572","id":"PMC_24962572","title":"G protein-coupled receptor 30 (GPR30) forms a plasma membrane complex with membrane-associated guanylate kinases (MAGUKs) and protein kinase A-anchoring protein 5 (AKAP5) that constitutively inhibits cAMP production.","date":"2014","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24962572","citation_count":54,"is_preprint":false},{"pmid":"20147557","id":"PMC_20147557","title":"Ca2+/calmodulin disrupts AKAP79/150 interactions with KCNQ (M-Type) K+ channels.","date":"2010","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/20147557","citation_count":53,"is_preprint":false},{"pmid":"21562284","id":"PMC_21562284","title":"AKAP79/150 signal complexes in G-protein modulation of neuronal ion channels.","date":"2011","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/21562284","citation_count":50,"is_preprint":false},{"pmid":"33941685","id":"PMC_33941685","title":"The N terminus of Orai1 couples to the AKAP79 signaling complex to drive NFAT1 activation by local Ca2+ entry.","date":"2021","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/33941685","citation_count":49,"is_preprint":false},{"pmid":"29196604","id":"PMC_29196604","title":"CaMKII regulates the depalmitoylation and synaptic removal of the scaffold protein AKAP79/150 to mediate structural long-term depression.","date":"2017","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29196604","citation_count":46,"is_preprint":false},{"pmid":"21273417","id":"PMC_21273417","title":"AKAP79/150 impacts intrinsic excitability of hippocampal neurons through phospho-regulation of A-type K+ channel trafficking.","date":"2011","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/21273417","citation_count":46,"is_preprint":false},{"pmid":"18305116","id":"PMC_18305116","title":"AKAP79 selectively enhances protein kinase C regulation of GluR1 at a Ca2+-calmodulin-dependent protein kinase II/protein kinase C site.","date":"2008","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/18305116","citation_count":46,"is_preprint":false},{"pmid":"19091974","id":"PMC_19091974","title":"Stable membrane expression of postsynaptic CaV1.2 calcium channel clusters is independent of interactions with AKAP79/150 and PDZ proteins.","date":"2008","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/19091974","citation_count":46,"is_preprint":false},{"pmid":"15458848","id":"PMC_15458848","title":"Modulation of dopamine mediated phosphorylation of AMPA receptors by PSD-95 and AKAP79/150.","date":"2004","source":"Neuropharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/15458848","citation_count":46,"is_preprint":false},{"pmid":"19858198","id":"PMC_19858198","title":"Ca2+/calmodulin-dependent protein kinase II binds to and phosphorylates a specific SAP97 splice variant to disrupt association with AKAP79/150 and modulate alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-type glutamate receptor (AMPAR) activity.","date":"2009","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19858198","citation_count":45,"is_preprint":false},{"pmid":"24121510","id":"PMC_24121510","title":"Role of AKAP79/150 protein in β1-adrenergic receptor trafficking and signaling in mammalian cells.","date":"2013","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24121510","citation_count":44,"is_preprint":false},{"pmid":"23699529","id":"PMC_23699529","title":"Mapping the binding site of TRPV1 on AKAP79: implications for inflammatory hyperalgesia.","date":"2013","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/23699529","citation_count":40,"is_preprint":false},{"pmid":"33082339","id":"PMC_33082339","title":"AKAP5 complex facilitates purinergic modulation of vascular L-type Ca2+ channel CaV1.2.","date":"2020","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/33082339","citation_count":38,"is_preprint":false},{"pmid":"32601188","id":"PMC_32601188","title":"STIM2 targets Orai1/STIM1 to the AKAP79 signaling complex and confers coupling of Ca2+ entry with NFAT1 activation.","date":"2020","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/32601188","citation_count":38,"is_preprint":false},{"pmid":"16442664","id":"PMC_16442664","title":"G-Protein-coupled receptor-associated A-kinase anchoring proteins: AKAP79 and AKAP250 (gravin).","date":"2006","source":"European journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/16442664","citation_count":37,"is_preprint":false},{"pmid":"16246112","id":"PMC_16246112","title":"Beta-arrestin-recruited phosphodiesterase-4 desensitizes the AKAP79/PKA-mediated switching of beta2-adrenoceptor signalling to activation of ERK.","date":"2005","source":"Biochemical Society transactions","url":"https://pubmed.ncbi.nlm.nih.gov/16246112","citation_count":35,"is_preprint":false},{"pmid":"19055733","id":"PMC_19055733","title":"G-protein-coupled receptor-associated A-kinase anchoring proteins AKAP5 and AKAP12: differential signaling to MAPK and GPCR recycling.","date":"2008","source":"Journal of molecular signaling","url":"https://pubmed.ncbi.nlm.nih.gov/19055733","citation_count":33,"is_preprint":false},{"pmid":"12938160","id":"PMC_12938160","title":"Identification of an IQGAP1/AKAP79 complex in beta-cells.","date":"2003","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12938160","citation_count":32,"is_preprint":false},{"pmid":"31091162","id":"PMC_31091162","title":"AKAP79/150 recruits the transcription factor NFAT to regulate signaling to the nucleus by neuronal L-type Ca2+ channels.","date":"2019","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/31091162","citation_count":32,"is_preprint":false},{"pmid":"10762347","id":"PMC_10762347","title":"Localization of the A kinase anchoring protein AKAP79 in the human hippocampus.","date":"2000","source":"The European journal of neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/10762347","citation_count":31,"is_preprint":false},{"pmid":"25225170","id":"PMC_25225170","title":"Carvedilol reverses cardiac insufficiency in AKAP5 knockout mice by normalizing the activities of calcineurin and CaMKII.","date":"2014","source":"Cardiovascular research","url":"https://pubmed.ncbi.nlm.nih.gov/25225170","citation_count":30,"is_preprint":false},{"pmid":"22976297","id":"PMC_22976297","title":"A key phosphorylation site in AC8 mediates regulation of Ca(2+)-dependent cAMP dynamics by an AC8-AKAP79-PKA signalling complex.","date":"2012","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/22976297","citation_count":30,"is_preprint":false},{"pmid":"23889134","id":"PMC_23889134","title":"AKAP79, PKC, PKA and PDE4 participate in a Gq-linked muscarinic receptor and adenylate cyclase 2 cAMP signalling complex.","date":"2013","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/23889134","citation_count":27,"is_preprint":false},{"pmid":"21156788","id":"PMC_21156788","title":"Ca2+/calmodulin-dependent protein kinase II inhibitors disrupt AKAP79-dependent PKC signaling to GluA1 AMPA receptors.","date":"2010","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21156788","citation_count":24,"is_preprint":false},{"pmid":"28967377","id":"PMC_28967377","title":"Intrinsic disorder within AKAP79 fine-tunes anchored phosphatase activity toward substrates and drug sensitivity.","date":"2017","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/28967377","citation_count":24,"is_preprint":false},{"pmid":"25504574","id":"PMC_25504574","title":"Anchoring protein AKAP79-mediated PKA phosphorylation of STIM1 determines selective activation of the ARC channel, a store-independent Orai channel.","date":"2015","source":"The Journal of physiology","url":"https://pubmed.ncbi.nlm.nih.gov/25504574","citation_count":24,"is_preprint":false},{"pmid":"22232585","id":"PMC_22232585","title":"A Potential Role for a Genetic Variation of AKAP5 in Human Aggression and Anger Control.","date":"2011","source":"Frontiers in human neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/22232585","citation_count":23,"is_preprint":false},{"pmid":"18950703","id":"PMC_18950703","title":"G-protein-coupled receptor-associated A-kinase anchoring proteins AKAP5 and AKAP12: differential trafficking and distribution.","date":"2008","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/18950703","citation_count":19,"is_preprint":false},{"pmid":"15914039","id":"PMC_15914039","title":"Differential expression of protein kinase A, AKAP79, and PP2B in pregnant human myometrial membranes prior to and during labor.","date":"2005","source":"Journal of the Society for Gynecologic Investigation","url":"https://pubmed.ncbi.nlm.nih.gov/15914039","citation_count":18,"is_preprint":false},{"pmid":"21561082","id":"PMC_21561082","title":"Identification of AKAP79 as a protein phosphatase 1 catalytic binding protein.","date":"2011","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21561082","citation_count":18,"is_preprint":false},{"pmid":"34612814","id":"PMC_34612814","title":"AKAP79 enables calcineurin to directly suppress protein kinase A activity.","date":"2021","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/34612814","citation_count":17,"is_preprint":false},{"pmid":"34458850","id":"PMC_34458850","title":"AKAP79 Orchestrates a Cyclic AMP Signalosome Adjacent to Orai1 Ca2+ Channels.","date":"2021","source":"Function (Oxford, England)","url":"https://pubmed.ncbi.nlm.nih.gov/34458850","citation_count":15,"is_preprint":false},{"pmid":"26170881","id":"PMC_26170881","title":"Curcumin Protects Neurons from Glutamate-Induced Excitotoxicity by Membrane Anchored AKAP79-PKA Interaction Network.","date":"2015","source":"Evidence-based complementary and alternative medicine : eCAM","url":"https://pubmed.ncbi.nlm.nih.gov/26170881","citation_count":15,"is_preprint":false},{"pmid":"21831305","id":"PMC_21831305","title":"AKAP12 and AKAP5 form higher-order hetero-oligomers.","date":"2011","source":"Journal of molecular signaling","url":"https://pubmed.ncbi.nlm.nih.gov/21831305","citation_count":14,"is_preprint":false},{"pmid":"33617875","id":"PMC_33617875","title":"AKAP79/150 coordinates leptin-induced PKA signaling to regulate KATP channel trafficking in pancreatic β-cells.","date":"2021","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/33617875","citation_count":14,"is_preprint":false},{"pmid":"30737285","id":"PMC_30737285","title":"Preferential generation of Ca2+-permeable AMPA receptors by AKAP79-anchored protein kinase C proceeds via GluA1 subunit phosphorylation at Ser-831.","date":"2019","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30737285","citation_count":14,"is_preprint":false},{"pmid":"22693956","id":"PMC_22693956","title":"AKAP79/150 interacts with the neuronal calcium-binding protein caldendrin.","date":"2012","source":"Journal of neurochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22693956","citation_count":14,"is_preprint":false},{"pmid":"22677788","id":"PMC_22677788","title":"AKAP79 modulation of L-type channels involves disruption of intramolecular interactions in the CaV1.2 subunit.","date":"2012","source":"Channels (Austin, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/22677788","citation_count":14,"is_preprint":false},{"pmid":"25653013","id":"PMC_25653013","title":"Mechanisms and dynamics of AKAP79/150-orchestrated multi-protein signalling complexes in brain and peripheral nerve.","date":"2015","source":"The Journal of physiology","url":"https://pubmed.ncbi.nlm.nih.gov/25653013","citation_count":13,"is_preprint":false},{"pmid":"21554706","id":"PMC_21554706","title":"AKAP5 and AKAP12 Form Homo-oligomers.","date":"2011","source":"Journal of molecular signaling","url":"https://pubmed.ncbi.nlm.nih.gov/21554706","citation_count":12,"is_preprint":false},{"pmid":"29976855","id":"PMC_29976855","title":"Targeting FRET-Based Reporters for cAMP and PKA Activity Using AKAP79.","date":"2018","source":"Sensors (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/29976855","citation_count":12,"is_preprint":false},{"pmid":"23462372","id":"PMC_23462372","title":"Increased density of AKAP5-expressing neurons in the anterior cingulate cortex of subjects with bipolar disorder.","date":"2013","source":"Journal of psychiatric research","url":"https://pubmed.ncbi.nlm.nih.gov/23462372","citation_count":11,"is_preprint":false},{"pmid":"36317924","id":"PMC_36317924","title":"Nuanced Interactions between AKAP79 and STIM1 with Orai1 Ca2+ Channels at Endoplasmic Reticulum-Plasma Membrane Junctions Sustain NFAT Activation.","date":"2022","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/36317924","citation_count":9,"is_preprint":false},{"pmid":"20164376","id":"PMC_20164376","title":"The contribution of AKAP5 in amylase secretion from mouse parotid acini.","date":"2010","source":"American journal of physiology. Cell physiology","url":"https://pubmed.ncbi.nlm.nih.gov/20164376","citation_count":9,"is_preprint":false},{"pmid":"22996592","id":"PMC_22996592","title":"Kinetic and mechanistic differences in the interactions between caldendrin and calmodulin with AKAP79 suggest different roles in synaptic function.","date":"2012","source":"Journal of molecular recognition : JMR","url":"https://pubmed.ncbi.nlm.nih.gov/22996592","citation_count":8,"is_preprint":false},{"pmid":"14519485","id":"PMC_14519485","title":"Expression and intracellular localization of protein phosphatases 2A and 2B, protein kinase a, A-Kinase anchoring protein (AKAP79), and binding of the regulatory (RII) subunit of protein kinase a to AKAP79 in human myometrium.","date":"2003","source":"Journal of the Society for Gynecologic Investigation","url":"https://pubmed.ncbi.nlm.nih.gov/14519485","citation_count":7,"is_preprint":false},{"pmid":"26713362","id":"PMC_26713362","title":"Modified sympathetic nerve regulation in AKAP5-null mice.","date":"2015","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/26713362","citation_count":6,"is_preprint":false},{"pmid":"11275698","id":"PMC_11275698","title":"Development-related expression of AKAP79 in the striatal compartments of the human brain.","date":"2001","source":"Cells, tissues, organs","url":"https://pubmed.ncbi.nlm.nih.gov/11275698","citation_count":5,"is_preprint":false},{"pmid":"24926793","id":"PMC_24926793","title":"AKAP5 keeps L-type channels and NFAT on their toes.","date":"2014","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/24926793","citation_count":5,"is_preprint":false},{"pmid":"25789584","id":"PMC_25789584","title":"AKAP5 signaling complexes: focal points and functional properties.","date":"2015","source":"Neuro endocrinology letters","url":"https://pubmed.ncbi.nlm.nih.gov/25789584","citation_count":4,"is_preprint":false},{"pmid":"22583680","id":"PMC_22583680","title":"\"Shaping\" of cell signaling via AKAP-tethered PDE4D: Probing with AKAR2-AKAP5 biosensor.","date":"2012","source":"Journal of molecular signaling","url":"https://pubmed.ncbi.nlm.nih.gov/22583680","citation_count":4,"is_preprint":false},{"pmid":"16460836","id":"PMC_16460836","title":"Contextual utilization of enzymes in discrete AKAP79/150 signalling complexes.","date":"2006","source":"European journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/16460836","citation_count":4,"is_preprint":false},{"pmid":"32173522","id":"PMC_32173522","title":"AKAP5 anchors PKA to enhance regulation of the HERG channel.","date":"2020","source":"The international journal of biochemistry & cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/32173522","citation_count":3,"is_preprint":false},{"pmid":"31310785","id":"PMC_31310785","title":"Maturation of thalamocortical synapses in the somatosensory cortex depends on neocortical AKAP5 expression.","date":"2019","source":"Neuroscience letters","url":"https://pubmed.ncbi.nlm.nih.gov/31310785","citation_count":3,"is_preprint":false},{"pmid":"38798824","id":"PMC_38798824","title":"Akap5 links synaptic dysfunction to neuroinflammatory signaling in a mouse model of infantile neuronal ceroid lipofuscinosis.","date":"2024","source":"Frontiers in synaptic neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/38798824","citation_count":3,"is_preprint":false},{"pmid":"15003529","id":"PMC_15003529","title":"AKAP79 increases the functional expression of skeletal muscle Ca2+ channels in Xenopus oocytes.","date":"2004","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/15003529","citation_count":3,"is_preprint":false},{"pmid":"36238650","id":"PMC_36238650","title":"Metoprolol Mitigates Ischemic Heart Remodeling and Fibrosis by Increasing the Expression of AKAP5 in Ischemic Heart.","date":"2022","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/36238650","citation_count":2,"is_preprint":false},{"pmid":"39864527","id":"PMC_39864527","title":"Unveiling the significance of AKAP79/150 in the nervous system disorders: An emerging opportunity for future therapies?","date":"2025","source":"Neurobiology of disease","url":"https://pubmed.ncbi.nlm.nih.gov/39864527","citation_count":1,"is_preprint":false},{"pmid":"41640285","id":"PMC_41640285","title":"Maternal exercise improves vascular function in hypertensive offspring via A-kinase anchoring protein 150 gene (Akap5) epigenetic modifications.","date":"2026","source":"British journal of pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/41640285","citation_count":0,"is_preprint":false},{"pmid":"37810553","id":"PMC_37810553","title":"Retracted: Metoprolol Mitigates Ischemic Heart Remodeling and Fibrosis by Increasing the Expression of AKAP5 in Ischemic Heart.","date":"2023","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/37810553","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.10.27.616460","title":"Targeting ADORA-PDE10 cAMP Microdomain: A Novel Therapeutic Approach for Pulmonary Hypertension","date":"2024-10-27","source":"bioRxiv","url":"https://doi.org/10.1101/2024.10.27.616460","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":46743,"output_tokens":13076,"usd":0.168184,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":24796,"output_tokens":6167,"usd":0.139077,"stage2_stop_reason":"end_turn"},"total_usd":0.307261,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1996,\n      \"finding\": \"AKAP79 functions as a scaffold protein that simultaneously binds three signaling enzymes—PKA, calcineurin (PP2B), and PKC—at distinct binding sites, coordinating their localization in mammalian neurons.\",\n      \"method\": \"Deletion analysis and binding studies (co-immunoprecipitation, subcellular co-distribution by immunofluorescence)\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal binding studies with deletion analysis, replicated across multiple subsequent labs\",\n      \"pmids\": [\"8599116\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1993,\n      \"finding\": \"AKAP79 (AKAP75) contains two distinct non-contiguous N-terminal domains (residues 27–48 and 77–91) required for intracellular membrane/cytoskeletal targeting, and a C-terminal RII-binding (tethering) domain mapped to residues 392–413 where hydrophobic residues are essential for high-affinity PKA regulatory subunit binding.\",\n      \"method\": \"Deletion and scanning mutagenesis with subcellular fractionation and RII binding assays in HEK293 cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — systematic mutagenesis with in vitro binding assays defining domain boundaries\",\n      \"pmids\": [\"8509414\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"Ca²⁺/calmodulin binds AKAP79 with high affinity (KD ~28 nM) and competes with PKC for the same N-terminal region (residues 31–52), releasing inhibited PKC from the AKAP79 complex and increasing PKC activity at postsynaptic densities.\",\n      \"method\": \"Calmodulin binding assays, co-immunoprecipitation, PKC activity assays, immunofluorescence in hippocampal neurons\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro binding with KD measurement, functional PKC activity assay, cellular co-immunoprecipitation\",\n      \"pmids\": [\"9202019\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"AKAP79 membrane targeting is mediated by three basic/hydrophobic N-terminal regions (A: residues 31–52; B: 76–101; C: 116–145) that bind acidic phospholipids including PtdIns(4,5)P2; this binding is regulated by phosphorylation and Ca²⁺/calmodulin, and PKC or calmodulin activation releases AKAP79 from membrane fractions.\",\n      \"method\": \"GFP-based fluorescence imaging of deletion mutants, membrane vesicle lipid-binding assays, subcellular fractionation, phosphorylation assays in HEK293 cells and cortical neurons\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (imaging, lipid binding, fractionation, phosphorylation) in a single study\",\n      \"pmids\": [\"9545238\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"AKAP79 binds calcineurin A through residues 108–280 on AKAP79 and residues 30–98 and 311–336 on calcineurin A, independently of calcineurin B and at a site distinct from immunophilin-binding regions; overexpression of AKAP79 inhibits calcineurin-mediated NFAT dephosphorylation and activation in intact cells.\",\n      \"method\": \"Co-immunoprecipitation, truncation mapping, NFAT reporter assay in transfected cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — domain mapping with co-IP plus functional NFAT reporter assay in cells\",\n      \"pmids\": [\"9765270\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"AKAP79 binds and inhibits the conserved catalytic core of PKC (multiple conventional, novel, and atypical isoforms) through a mechanism involving the pseudosubstrate displacement by residues 31–52 (specifically Arg39 and Arg40); lipid activators or kinase activation are not required for the association.\",\n      \"method\": \"In vitro binding and kinase activity assays, endoproteinase proteolysis, mutagenesis of AKAP79 and PKCβII, co-immunoprecipitation and immunofluorescence in hippocampal neurons\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with mutagenesis and multiple PKC isoforms tested\",\n      \"pmids\": [\"10510312\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"AKAP79 regulates GRK2 membrane recruitment and β2AR phosphorylation by anchoring PKA, which directly phosphorylates GRK2 on Ser685, increasing Gβγ binding to GRK2 and enhancing GRK2 translocation to agonist-occupied receptor; S685A mutation or dominant-negative AKAP79 reduces GRK2-mediated receptor phosphorylation and internalization.\",\n      \"method\": \"Mutagenesis of GRK2 (S685A), dominant-negative AKAP79 expression, phosphorylation assays, receptor internalization assays in HEK293 cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — site-directed mutagenesis plus functional phosphorylation and internalization assays\",\n      \"pmids\": [\"11278469\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"AKAP79 directly associates with the inwardly rectifying potassium channel Kir2.1 via both N- and C-terminal intracellular domains of the channel, and this association enhances the channel's response to elevated intracellular cAMP.\",\n      \"method\": \"Co-immunoprecipitation from intact cells, GST pulldown with Kir2.1 intracellular domains, electrophysiology in transfected cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — GST pulldown plus co-IP plus functional electrophysiology, single lab\",\n      \"pmids\": [\"11287423\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"AKAP79 promotes basal PKA-mediated phosphorylation of GluR1 Ser845 and, through anchored PP2B and Ca²⁺ signaling, confers calcineurin-dependent downregulation of GluR1 receptor currents analogous to LTD; this requires PKA, Ser845, and PDZ-domain interaction between GluR1 and SAP97.\",\n      \"method\": \"Electrophysiology, co-immunoprecipitation, phosphorylation assays in HEK293 cells and hippocampal neurons\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal biochemistry plus functional electrophysiology with mutagenesis controls\",\n      \"pmids\": [\"11943807\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"AKAP79 directly regulates cell-surface expression (trafficking) of L-type calcium channels (CaV1.2) via a polyproline sequence in the channel II–III cytoplasmic loop, independently of PKA activity.\",\n      \"method\": \"Extracellular epitope tagging of CaV1.2, immunoassays, whole-cell and single-channel electrophysiology in transfected cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional imaging plus electrophysiology, single lab\",\n      \"pmids\": [\"12114507\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"The PP2B/calcineurin-binding site on AKAP79 is localized to residues 315–360; multiple determinants in this region bind directly to the calcineurin A subunit and inhibit phosphatase activity; peptides spanning residues 330–357 antagonize PP2B anchoring and attenuate PP2B-dependent downregulation of GluR1 currents.\",\n      \"method\": \"Cellular targeting assays with AKAP79 truncation/deletion mutants, in vitro binding and phosphatase activity assays, peptide antagonist electrophysiology in HEK293 cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — systematic mutagenesis with in vitro phosphatase assay plus functional electrophysiology\",\n      \"pmids\": [\"12354762\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"FRET microscopy in living cells directly demonstrated that PKA-RII and calcineurin A subunits simultaneously bind AKAP79 at the plasma membrane cytoskeleton within ~5 nm of each other, forming a ternary kinase-scaffold-phosphatase complex; AKAP79 also regulates membrane localization of SAP97.\",\n      \"method\": \"FRET microscopy (immunofluorescence and live-cell FRET), subcellular localization in transfected cells\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — FRET in live cells providing direct distance measurement of ternary complex\",\n      \"pmids\": [\"12507994\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"AKAP79 directly interacts with the C-terminal domain of IQGAP1, forming an IQGAP1/AKAP79 complex that co-purifies with PKA in β-cells, potentially linking cAMP/PKA signaling with Ca²⁺/CaM and GTPase pathways.\",\n      \"method\": \"cAMP affinity chromatography, co-immunoprecipitation, direct binding assay with IQGAP1 C-terminal domain\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — pulldown and co-IP, single lab, limited functional follow-up\",\n      \"pmids\": [\"12938160\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"AKAP79 is constitutively associated with the β2-adrenergic receptor and anchors PKA to mediate PKA-dependent phosphorylation of β2AR and switching of β2AR signaling to ERK activation via Gi; β-arrestin-recruited PDE4D5 desensitizes this AKAP79/PKA-mediated process.\",\n      \"method\": \"siRNA knockdown of specific PDE4 isoforms and AKAP79, co-immunoprecipitation, β2AR phosphorylation assays, ERK activation assays in HEK293B2 cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — isoform-selective siRNA knockdown with multiple orthogonal functional readouts\",\n      \"pmids\": [\"16030021\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"AKAP79-mediated targeting of PKA to the β1-adrenergic receptor promotes receptor recycling and functional resensitization; AKAP79, β1-AR, and PKA form a ternary complex at the β1-AR C-terminus; siRNA knockdown of AKAP79 prevents β1-AR recycling.\",\n      \"method\": \"siRNA knockdown, co-immunoprecipitation, FRET microscopy, PKA phosphorylation assays in HEK293 cells, SK-N-MC cells, and neonatal rat cortical neurons\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — siRNA rescue plus FRET plus phosphorylation assays, multiple cell types\",\n      \"pmids\": [\"16940053\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"AKAP79 forms a complex with SAP97 at the PDZ-binding sequence (ESKV) at the β1-AR C-terminus, assembling a receptosome that targets PKA to β1-AR for Ser312 phosphorylation in the third intracellular loop, which is required for receptor recycling.\",\n      \"method\": \"Co-immunoprecipitation, domain deletion mutants, PKA phosphorylation assays, receptor trafficking assays in HEK293 cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with mutagenesis and functional trafficking readout, single lab\",\n      \"pmids\": [\"17170109\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"AKAP79/150 directly interacts with CaV1.2 and co-targets PKA and calcineurin to L-type channels, conferring bidirectional regulation of L-type current; anchored calcineurin dominantly suppresses PKA enhancement of the channel; AKAP79/150-anchored calcineurin is required for NFATc4 activation by local Ca²⁺ influx through L-type channels.\",\n      \"method\": \"Co-immunoprecipitation, electrophysiology in HEK293 cells and hippocampal neurons, NFAT reporter assays, dominant-negative AKAP constructs\",\n      \"journal\": \"Neuron\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct binding plus electrophysiology plus transcription factor activation assay in two cell systems\",\n      \"pmids\": [\"17640527\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"AKAP79/150 forms a complex with TRPV1, PKA, PKC, and calcineurin via a critical region in the TRPV1 C-terminus; disrupting this binding abrogates TRPV1 sensitization by bradykinin and PGE2, demonstrating that AKAP79/150 is a final common element for heat hyperalgesia.\",\n      \"method\": \"Co-immunoprecipitation, deletion mapping of TRPV1 C-terminus, TRPV1 electrophysiology in transfected cells and neurons, in vivo hyperalgesia assays\",\n      \"journal\": \"Neuron\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — domain mapping, co-IP, functional electrophysiology, and in vivo behavioral assay\",\n      \"pmids\": [\"18701070\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"AKAP79 selectively enhances PKC-mediated phosphorylation of GluR1 Ser831 by localizing PKC near the receptor via SAP97, shifting the PKC dose-dependence ~20-fold so that low PKC concentrations are as effective as much higher CaMKII concentrations.\",\n      \"method\": \"Biochemical phosphorylation assays, electrophysiology of GluR1 currents in transfected cells with AKAP79 constructs\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical and electrophysiological assays, single lab\",\n      \"pmids\": [\"18305116\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"AKAP79 associates with multiple adenylyl cyclase (AC) isoforms (AC5, AC6, AC9) via their N-terminal regions and residues 77–108 of AKAP79; this interaction places AC5/6 in proximity to synaptic AMPA receptors; loss of AKAP150 in mice decreases AMPA receptor-associated AC activity.\",\n      \"method\": \"Co-immunoprecipitation, FRET (intensity- and lifetime-based) in living cells, AC activity assays in brain extracts from AKAP150 KO mice, domain deletion mapping\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — FRET in living cells plus biochemical assays plus mouse KO validation\",\n      \"pmids\": [\"20231277\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"AKAP5 (AKAP79/150) organizes a caveolin-3-associated signaling complex in cardiac T-tubules comprising adenylyl cyclase 5/6, PKA, calcineurin, and a subpopulation of CaV1.2 channels; only this caveolin-3-associated CaV1.2 subpopulation is phosphorylated by PKA upon sympathetic stimulation; AKAP5 KO disrupts this complex, preventing normal calcium transients and PKA-dependent phosphorylation of ryanodine receptors and phospholamban.\",\n      \"method\": \"AKAP5 knockout mice, calcium imaging, electrophysiology, co-immunoprecipitation, PKA phosphorylation assays in cardiomyocytes\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO with multiple orthogonal biochemical and functional readouts\",\n      \"pmids\": [\"20671242\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"AKAP79 alters the cellular pharmacology of anchored PKC, protecting it from certain ATP-competitive inhibitors; AKAP79-anchored PKC synchronizes muscarinic agonist-induced phosphorylation and KCNQ2 M-channel inhibition to optimize neuronal excitability.\",\n      \"method\": \"Dual fluorescent imaging/patch-clamp technique with CKAR kinase activity reporter, pharmacological profiling, electrophysiology in neurons\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — simultaneous live imaging and electrophysiology with pharmacological dissection, rigorous controls\",\n      \"pmids\": [\"20188672\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"AKAP79/150 directly interacts with AC8, limiting AC8 sensitivity to intracellular Ca²⁺; this interaction was observed in HEK293 cells, pancreatic insulin-secreting cells, and hippocampal neurons.\",\n      \"method\": \"Co-immunoprecipitation, high-resolution live-cell imaging in multiple cell types endogenously expressing both proteins\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP in multiple physiologically relevant cell types, single lab\",\n      \"pmids\": [\"20410303\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"AKAP5 knockout in mice delocalizes PKA from hippocampus and striatum (redistributing it to dendritic shafts via MAP2 binding); the D36 mutant lacking only the PKA binding domain produces more severe electrophysiological and behavioral deficits than complete KO, indicating that targeting calcineurin or other binding partners without the balancing PKA disrupts synaptic plasticity.\",\n      \"method\": \"Genetic KO and knockin (D36) mouse lines, immunofluorescence, co-immunoprecipitation, electrophysiology, behavioral tests\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with two distinct mutant lines plus multiple biochemical and functional readouts\",\n      \"pmids\": [\"20428246\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Ca²⁺/calmodulin disrupts AKAP79/150 interaction with KCNQ2–5 (but not KCNQ1) channels as shown by TIRF/FRET; this disruption prevents AKAP79-mediated sensitization of KCNQ2/3 channels to muscarinic inhibition, while PIP2 depletion does not affect AKAP79 membrane localization.\",\n      \"method\": \"TIRF/FRET microscopy, perforated patch-clamp electrophysiology, dominant-negative calmodulin, cotransfection in CHO cells\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — TIRF/FRET with electrophysiology and multiple molecular tools (WT vs DN CaM, PIP2 phosphatase)\",\n      \"pmids\": [\"20147557\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"AKAP79 dimerizes through K328-K328 and K333-K333 cross-links; the reconstituted AKAP79-PP2B-RII-CaM complex has a molecular mass of ~466 kDa consisting of dimeric AKAP79 coordinating two RII homodimers, four PP2B heterodimers, and two calmodulin molecules; Ca²⁺/CaM binding activates anchored phosphatases by generating a second interface.\",\n      \"method\": \"Native mass spectrometry, chemical cross-linking, in vitro reconstitution of quaternary complex, quantitative biochemical analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with native MS and cross-linking providing stoichiometry and architecture\",\n      \"pmids\": [\"21464287\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"AKAP79/150-anchored PKA controls Kv4.2 surface expression; the C-terminal domain of Kv4.2 interacts with an internal region of AKAP79/150 overlapping its MAGUK-binding domain; disrupting PKA anchoring decreases neuronal excitability while preventing calcineurin-mediated dephosphorylation increases excitability.\",\n      \"method\": \"Co-immunoprecipitation, pulldown, AKAP79/150 KO neurons, PKA anchoring disruption (Ht31 peptide), electrophysiology in hippocampal neurons\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — binding domain mapping plus functional electrophysiology, single lab\",\n      \"pmids\": [\"21273417\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"AKAP79 is identified as a novel PP1 regulatory subunit; it directly binds PP1 catalytic subunit through a consensus FxxR/KxR/K motif in residues 1–44 and a second domain in residues 150–250; AKAP79 inhibits PP1 activity (IC50 ~811 nM) in a substrate-dependent manner but does not inhibit PP1 dephosphorylation of phospho-PSD-95.\",\n      \"method\": \"Co-immunoprecipitation from rat brain, pulldown with purified proteins, phosphatase activity assays, surface plasmon resonance, peptide mapping\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro assays with purified proteins plus SPR, single lab\",\n      \"pmids\": [\"21561082\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Palmitoylation of AKAP79 at two N-terminal cysteines targets it to lipid rafts; mutation of these cysteines excludes AKAP79 from rafts, alters membrane diffusion behavior, and abolishes AKAP79-dependent regulation of SOCE-stimulated AC8 activity and PKA-dependent phosphorylation of raft proteins.\",\n      \"method\": \"Pharmacological palmitoylation inhibition, site-directed mutagenesis of Cys residues, lipid raft fractionation, FRAP analysis, AC8 activity assays in HEK293 cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — mutagenesis plus biochemical fractionation plus functional AC8 assay, multiple orthogonal methods\",\n      \"pmids\": [\"21771783\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"AKAP79/150-mediated PKC anchoring is specifically required for muscarinic M1 (and angiotensin II) receptor suppression of M-type (KCNQ) K⁺ currents in SCG neurons, but not for bradykinin or purinergic suppression; FRET showed strong AKAP79 association with M1 and AT1 receptors and KCNQ2/3, but weak association with P2Y6 or B2 receptors.\",\n      \"method\": \"AKAP150 KO mice, transfection of ΔA-AKAP79 (no PKC binding), FRET under TIRF microscopy, perforated patch-clamp in SCG neurons\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO complemented by domain-specific mutant rescue plus FRET plus electrophysiology\",\n      \"pmids\": [\"21562284\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"The IAIIIT anchoring motif in AKAP79 (residues forming a short linear motif) binds the same surface of calcineurin as the PxIxIT recognition peptide of NFAT but with higher affinity; increasing calcineurin-AKAP affinity paradoxically impairs NFAT activation by slowing calcineurin release and sequestering it as 'decoy' sites.\",\n      \"method\": \"Structural binding analysis, mutagenesis of AKAP79 anchoring sequence, NFAT activation assays, calcineurin recruitment measurements\",\n      \"journal\": \"Nature structural & molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — structural binding analysis plus mutagenesis plus functional NFAT assay revealing mechanistic balance\",\n      \"pmids\": [\"22343722\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"AKAP79/150-anchored calcineurin and L-type Ca²⁺ channel activation drive KCNQ2/3 transcriptional upregulation via NFAT in hippocampal neurons; AKAP150 KO mice fail to upregulate KCNQ2/3 transcription after drug-induced seizures, indicating a negative feedback mechanism on neuronal excitability.\",\n      \"method\": \"Neuronal activity manipulation, NFAT reporter assays, AKAP150 KO mice, qRT-PCR of KCNQ2/3 mRNA in seizure model\",\n      \"journal\": \"Neuron\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO in physiological seizure model plus mechanistic reporter assays\",\n      \"pmids\": [\"23259949\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"AKAP79 interacts with caldendrin (a neuronal Ca²⁺-binding protein) at a site overlapping the calmodulin-binding region; caldendrin competes with calmodulin for binding to AKAP79 with similar affinity (KD ~20 nM vs ~30 nM for CaM), but through an induced-fit mechanism with a slow rearrangement step and Ca²⁺-independent binding component.\",\n      \"method\": \"Pulldown experiments, surface plasmon resonance biosensor kinetic analysis\",\n      \"journal\": \"Journal of neurochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — SPR with kinetic modeling plus pulldown, single lab\",\n      \"pmids\": [\"22693956\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"AKAP79 modulation of CaV1.2 membrane expression occurs through disruption of an autoinhibitory intramolecular interaction between the channel II–III linker and distal C-terminus; AKAP79 directly interacts with the distal CaV1.2 C-terminus, competing with its association to the II–III linker.\",\n      \"method\": \"Mutagenesis of polyproline domains, co-immunoprecipitation, channel trafficking assays in transfected cells\",\n      \"journal\": \"Channels\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mutagenesis plus co-IP plus functional channel expression assay, single lab\",\n      \"pmids\": [\"22677788\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"PKA recruited to AC8 via AKAP79 phosphorylates AC8 at Ser112, providing a negative feedback mechanism that reduces the on-rate of cAMP production during Ca²⁺ oscillations.\",\n      \"method\": \"Site-directed mutagenesis of AC8 (S112A), FRET-based cAMP measurements during Ca²⁺ oscillations, co-immunoprecipitation\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — mutagenesis of substrate site plus live-cell FRET functional assay\",\n      \"pmids\": [\"22976297\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"The AKAP79 region between amino acids 326–336 mediates binding to TRPV1; a TAT-conjugated peptide mimicking this domain inhibits TRPV1 sensitization in vitro and inflammatory hyperalgesia in vivo without affecting basal pain thresholds.\",\n      \"method\": \"FRET, co-immunoprecipitation, TRPV1 membrane trafficking assays, in vivo hyperalgesia assay with TAT-peptide\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — domain mapping with FRET/co-IP plus in vivo functional validation\",\n      \"pmids\": [\"23699529\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"AKAP5 (AKAP79/150) anchors adenylyl cyclase (AC) to postsynaptic sites and this AC anchoring is required for β-adrenergic stimulation-induced phosphorylation of GluA1 Ser845 and for theta-frequency-induced LTP; AC anchoring (disrupted in AKAP5 KO) is more critical than PKA anchoring alone (disrupted in D36 mice) for this process.\",\n      \"method\": \"AKAP5 KO and D36 (PKA-binding-deleted) knock-in mice, phosphorylation assays, LTP electrophysiology in acute forebrain slices\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — two distinct genetic mouse models with biochemical and electrophysiological readouts\",\n      \"pmids\": [\"23649627\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"AKAP79 recruits PKC to activate AC2 and generates localized cAMP upon Gq-coupled muscarinic receptor stimulation; PKA anchored to AKAP79 activates PDE4 to degrade this cAMP; calcineurin anchored to AKAP79 is not involved in this pathway.\",\n      \"method\": \"Live-cell cAMP imaging, siRNA knockdown of AKAP79 components, pharmacological dissection in HEK293 cells\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — live-cell imaging plus siRNA dissection, single lab\",\n      \"pmids\": [\"23889134\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"AKAP5 scaffold is required for PKA-GluA1 coupling during homeostatic plasticity; knockdown of AKAP5 blocks synaptic scaling up, which requires PKA-mediated phosphorylation of GluA1 S845; scaling down involves loss of PKA from synapses, and scaling up involves enhanced synaptic PKA activity regulated by AKAP5.\",\n      \"method\": \"AKAP5 RNAi knockdown, GluA1 S845 knockin mutant, phosphorylation assays, electrophysiology, surface AMPAR trafficking assays in neurons\",\n      \"journal\": \"Neuron\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — knockin mutation plus RNAi plus phosphorylation and trafficking assays with functional electrophysiology\",\n      \"pmids\": [\"25451194\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"AKAP5 controls calcineurin (CaN) and CaMKII activity in cardiac myocytes; loss of AKAP5 enhances CaN and CaMKII activities, interferes with β1-AR recycling through CaN binding to AKAP5, and leads to cardiac dilatation and dysfunction.\",\n      \"method\": \"AKAP5 KO mice, NFAT-luciferase reporter, echocardiography, biochemical assays, pharmacological rescue with carvedilol\",\n      \"journal\": \"Cardiovascular research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with functional cardiac readouts and mechanistic reporter assay, single lab\",\n      \"pmids\": [\"25225170\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"DHHC2 palmitoyl acyltransferase, resident in recycling endosomes, directly palmitoylates AKAP79/150 to regulate its targeting to recycling endosomes; DHHC2 knockdown disrupts recycling endosome exocytosis, dendritic spine enlargement, AKAP recruitment to spines, and LTP-stimulated AMPAR delivery; a palmitoylation-independent lipidated AKAP mutant rescues these deficits.\",\n      \"method\": \"RNAi knockdown of DHHC2, palmitoylation-independent AKAP rescue mutant, live imaging of spine exocytosis, electrophysiology in hippocampal neurons\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — RNAi with mechanistic rescue mutant plus multiple structural and functional plasticity readouts\",\n      \"pmids\": [\"25589740\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"AKAP79 constitutively associates with plasma membrane STIM1 and mediates PKA phosphorylation of STIM1 Thr389, which is necessary for activation of store-independent ARC channels but actually inhibits STIM1's ability to activate store-operated CRAC channels.\",\n      \"method\": \"AKAP79 knockdown, STIM1 T389 mutagenesis, ARC and CRAC channel electrophysiology, PKA activity assays\",\n      \"journal\": \"The Journal of physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — site-directed mutagenesis of substrate residue plus functional electrophysiology distinguishing two channel types\",\n      \"pmids\": [\"25504574\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"CaMKII mediates LTD-induced synaptic removal of AKAP79/150 by phosphorylating its N-terminal polybasic targeting domain (inhibiting F-actin association) and by promoting depalmitoylation at two N-terminal Cys residues; depalmitoylation (not phosphorylation per se) is required for AKAP79/150 spine removal and LTD-induced spine shrinkage; autonomous CaMKII activity preferentially phosphorylates AKAP79/150 compared with Ca²⁺/CaM-stimulated CaMKII.\",\n      \"method\": \"CaMKII inhibitors and constitutively active CaMKII, mutagenesis of palmitoylation sites and phosphorylation sites, live imaging of spine morphology, AKAP trafficking in hippocampal neurons during LTD\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple mutagenesis approaches plus pharmacological dissection plus live imaging with functional LTD readout\",\n      \"pmids\": [\"29196604\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Intrinsic disorder in AKAP79 allows conformational flexibility in PP2B engagement; the sole PP2B-anchoring determinant is a short linear motif (residues 337–343); Ca²⁺/calmodulin activation condenses diverse conformational variants into a uniform 178 Å population and engages a Leu-Lys-Ile-Pro sequence (residues 125–128) that occupies a PP2B binding pocket shared with cyclosporin, fine-tuning anchored phosphatase drug sensitivity.\",\n      \"method\": \"Negative-stain electron microscopy structural analysis, live-cell fluorescent activity sensor imaging, mutagenesis\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — EM structural analysis plus live-cell functional imaging plus mutagenesis in single study\",\n      \"pmids\": [\"28967377\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"AKAP79 directly interacts with NFAT via its C-terminal leucine-zipper (LZ) domain; disrupting this LZ interaction abolishes depolarization-stimulated NFAT signaling in hippocampal neurons while preserving the AKAP-LTCC interaction and LTCC function; AKAP79 thus recruits NFAT to the LTCC signaling complex to promote its activation by anchored calcineurin.\",\n      \"method\": \"RNAi knockdown of AKAP150 with human AKAP79 LZ-mutant replacement, FRET, Ca²⁺ imaging, electrophysiology, NFAT reporter, FRAP and FCS in hippocampal neurons\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — domain-specific mutant rescue plus FRET plus multiple functional assays in neurons\",\n      \"pmids\": [\"31091162\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"AKAP79-anchored PKC (not other AKAP79-signaling components) drives the appearance of Ca²⁺-permeable (GluA2-lacking) AMPARs primarily through GluA1 Ser831 phosphorylation; this generates CP-AMPARs under conditions where CI-AMPARs normally predominate.\",\n      \"method\": \"Electrophysiology (current-voltage relationships), GluA1 phosphosite mutagenesis, AKAP79 domain mutants in transfected cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mutagenesis plus electrophysiology, single lab\",\n      \"pmids\": [\"30737285\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"AKAP5 organizes a nanocomplex of P2Y11/P2Y11-like receptors, AC5, PKA, and CaV1.2 at the plasma membrane of arterial myocytes; disruption of AKAP5 function blocks glucose- and P2Y11 agonist-induced cAMP synthesis, CaV1.2 potentiation, vasoconstriction, and decreased blood flow.\",\n      \"method\": \"AKAP5 null mice and arterial myocytes, proximity ligation assay for nanocomplexes, cAMP assays, electrophysiology, vasoconstriction assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO plus proximity ligation plus multiple functional vascular readouts\",\n      \"pmids\": [\"33082339\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"STIM2 recruits Orai1/STIM1 to ER-PM junctions in response to ER-Ca²⁺ depletion, promoting assembly with AKAP79 to couple Orai1 channel function to NFAT1 activation; STIM2 knockdown attenuates NFAT1 activation and Orai1-AKAP79 assembly without substantially reducing Orai1/STIM1 clustering or global Ca²⁺ increases.\",\n      \"method\": \"STIM2 knockdown, STIM1ΔK mutant, co-immunoprecipitation, Ca²⁺ imaging, NFAT1 nuclear translocation assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockdown plus multiple mutant constructs plus biochemical and functional Ca²⁺/NFAT assays\",\n      \"pmids\": [\"32601188\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"AKAP79 enables calcineurin to dephosphorylate PKA type II regulatory subunits at a rate ~10-fold higher than without scaffolding; this allows calcineurin to suppress PKA activity by increasing catalytic subunit capture rate without altering cAMP levels, and this mechanism contributes to hippocampal LTD.\",\n      \"method\": \"In vitro reconstitution phosphatase assays, fluorescent PKA activity reporter (AKAR), kinetic modeling, hippocampal neuron LTD electrophysiology\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution plus live-cell PKA reporter plus kinetic modeling plus LTD electrophysiology\",\n      \"pmids\": [\"34612814\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"The N-terminus of Orai1 (but not Orai2, Orai3, or a shorter Orai1 isoform) directly interacts with AKAP79; NMR structural analysis reveals the AKAP-binding domain has a compact shape with proline-driven turns; this interaction is essential for NFAT1 activation by local Ca²⁺ entry and cytokine production.\",\n      \"method\": \"NMR structural analysis of AKAP79-binding domain, co-immunoprecipitation, domain deletion/truncation of Orai isoforms, NFAT1 reporter assays, cytokine production assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — NMR structure plus biochemical binding mapping plus multiple functional assays\",\n      \"pmids\": [\"33941685\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"AKAP79 coordinates PKA and PDE4 in a cAMP signaling nexus adjacent to Orai1; both PKA and PDE4 associate with AKAP79 and relocalize close to Orai1 after stimulation; in HEK293 cells, which lack functional Ca²⁺-activated adenylyl cyclases including AC8, Ca²⁺ entry through Orai1 does not increase cAMP levels despite AKAP79-Orai1 association.\",\n      \"method\": \"FRET-based cAMP sensors (AKAP79-CUTie), mass spectrometry, PCR, bulk cAMP and PKA activity measurements in HEK293 cells\",\n      \"journal\": \"Function\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — targeted FRET reporter plus mass spectrometry with multiple negative controls, single lab\",\n      \"pmids\": [\"34458850\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"AKAP79/150 coordinates leptin-induced PKA signaling at the cell membrane in pancreatic β-cells; leptin increases PKA activity at the membrane via NMDAR-CaMKKβ-AMPK signaling in an AKAP79/150-dependent manner; disrupting PP2B anchoring to AKAP79/150 elevates basal PKA signaling and increases surface KATP channels even without leptin.\",\n      \"method\": \"FRET-based PKA activity reporters, AKAP79/150 genetic knockdown and rescue, dominant-negative PP2B anchoring disruption, KATP channel trafficking assays in MIN6 β-cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — FRET reporter plus genetic knockdown rescue plus channel trafficking assay, single lab\",\n      \"pmids\": [\"33617875\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"A pre-assembled RXFP1-AKAP79-AC2-β-arrestin 2-PDE4D3 signalosome mediates sub-picomolar relaxin signaling; AC2 is functionally coupled to RXFP1 through AKAP79 binding to helix 8 of the receptor; PKA-activated PDE4D3 scaffolded via β-arrestin 2 tonically opposes AC2 activity.\",\n      \"method\": \"cAMP biosensors in single cells, co-immunoprecipitation, domain mapping (helix 8 of RXFP1)\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — live-cell cAMP imaging plus biochemical co-IP with domain identification, single lab\",\n      \"pmids\": [\"20664520\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The AKAP79-Orai1 interaction is considerably more transient than STIM1-Orai1; free AKAP79 (with calcineurin and NFAT1) can rapidly replace AKAP79 devoid of NFAT1 on Orai1 during continuous Ca²⁺ entry; Ca²⁺ nanodomains near Orai1 activate nearly the entire cytosolic NFAT1 pool, and recycling of inactive NFAT1 from cytoplasm to AKAP79 sustains excitation-transcription coupling.\",\n      \"method\": \"FRAP, FCS, co-immunoprecipitation, NFAT1 translocation assays, mathematical kinetic modeling\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — FRAP/FCS kinetic measurements plus mathematical model, single lab\",\n      \"pmids\": [\"36317924\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"AKAP79/150 (AKAP5) is a multivalent postsynaptic scaffold protein that simultaneously anchors PKA, calcineurin (PP2B), PKC, PP1, and adenylyl cyclase isoforms (AC5/6/8/9) at the plasma membrane via distinct binding domains; it targets these enzymes to ion channels (CaV1.2, TRPV1, KCNQ2-5, Kir2.1, Kv4.2, KATP, HERG), GPCRs (β1-AR, β2-AR, RXFP1, GPR30), and AMPA receptors through direct protein-protein interactions or scaffold intermediaries (SAP97, caveolin-3), thereby organizing compartmentalized phosphorylation/dephosphorylation cycles that bidirectionally regulate synaptic plasticity, neuronal excitability, channel trafficking, NFAT-dependent transcription, and cardiac β-adrenergic signaling; its membrane targeting is determined by lipid (PtdIns(4,5)P2) binding through polybasic N-terminal regions and palmitoylation at two N-terminal cysteines (mediated by DHHC2), while LTD-stimulus-induced CaMKII phosphorylation and depalmitoylation drive its synaptic removal.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"AKAP5 (AKAP79/150) is a multivalent membrane-targeted scaffold that simultaneously anchors the kinase PKA, the phosphatase calcineurin (PP2B), and PKC at distinct binding sites to coordinate compartmentalized phosphorylation/dephosphorylation cycles in neurons and other tissues [#0]. Systematic mutagenesis defined its modular architecture: a C-terminal RII (PKA)-binding helix [#1], an internal calcineurin-anchoring site centered on a short IAIIIT/LxxIP linear motif [#10, #30, #43], and an N-terminal region that both binds and inhibits the PKC catalytic core via a pseudosubstrate-like Arg39/Arg40 mechanism [#5]. The same N-terminal polybasic/hydrophobic regions target the scaffold to the membrane through PtdIns(4,5)P2 and acidic phospholipid binding [#3] and through DHHC2-mediated palmitoylation of two N-terminal cysteines that direct it to lipid rafts and recycling endosomes [#28, #40]; Ca2+/calmodulin and PKC activation release AKAP5 from membranes and from channel partners, and calmodulin competes directly with PKC for the N-terminal site [#2, #3, #24]. Native mass spectrometry and EM established that AKAP5 dimerizes to assemble a ~466 kDa quaternary complex in which Ca2+/CaM binding generates a second interface that activates anchored phosphatase, and that intrinsic disorder permits conformational flexibility in PP2B engagement [#25, #43]. Through these modules AKAP5 docks PKA, calcineurin, PKC, adenylyl cyclases (AC2/5/6/8/9) and PP1 onto ion channels and receptors—CaV1.2, TRPV1, KCNQ2-5, Kir2.1, Kv4.2, KATP, Orai1/STIM, and β1/β2-adrenergic, muscarinic, P2Y and RXFP1 GPCRs—to bidirectionally control channel activity, trafficking and receptor recycling [#16, #17, #24, #26, #46, #49, #52]. In neurons this organization governs AMPA receptor phosphorylation and trafficking underlying LTP, LTD and homeostatic synaptic scaling [#8, #36, #38, #48], and couples local Ca2+ entry through L-type channels and Orai1 to calcineurin-dependent NFAT transcription, including feedback upregulation of KCNQ channels [#16, #31, #44, #49]. In cardiac and arterial myocytes AKAP5 nucleates caveolin-3- and AC5-associated nanocomplexes that confer β-adrenergic and purinergic control of CaV1.2, calcium handling, and vasoconstriction, with knockout causing cardiac dilatation and dysfunction [#20, #39, #46]. AKAP5 also functions as a PKA-dependent regulatory node for GRK2 recruitment and β2AR desensitization [#6] and acts directly as a PP1 inhibitory subunit [#27].\",\n  \"teleology\": [\n    {\n      \"year\": 1993,\n      \"claim\": \"Established the modular domain organization that makes AKAP5 a targeting scaffold, separating membrane/cytoskeletal anchoring from PKA tethering.\",\n      \"evidence\": \"Deletion and scanning mutagenesis with subcellular fractionation and RII binding assays in HEK293 cells\",\n      \"pmids\": [\"8509414\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not address how the same protein engages calcineurin or PKC\", \"Membrane targets of the N-terminal domains not yet identified\"]\n    },\n    {\n      \"year\": 1996,\n      \"claim\": \"Defined the central concept that AKAP5 is a single scaffold binding three signaling enzymes (PKA, calcineurin, PKC) at distinct sites, enabling coordinated localization.\",\n      \"evidence\": \"Deletion analysis and binding studies with co-IP and immunofluorescence co-distribution in neurons\",\n      \"pmids\": [\"8599116\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether the three enzymes occupy one complex simultaneously not directly shown\", \"No functional readout of coordinated signaling\"]\n    },\n    {\n      \"year\": 1997,\n      \"claim\": \"Showed Ca2+/calmodulin acts as a regulatory switch on the scaffold by competing with PKC for the N-terminal site and releasing active PKC.\",\n      \"evidence\": \"Calmodulin binding assays with KD measurement, co-IP, PKC activity assays in hippocampal neurons\",\n      \"pmids\": [\"9202019\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological trigger of CaM competition in vivo not established\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"Resolved how AKAP5 reaches the membrane and how PKC inhibition works, linking lipid binding and pseudosubstrate displacement to regulated targeting.\",\n      \"evidence\": \"GFP imaging of mutants, lipid-binding/fractionation assays, in vitro PKC binding and kinase assays with mutagenesis (combined across studies)\",\n      \"pmids\": [\"9545238\", \"9765270\", \"10510312\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative in vivo contribution of lipid binding vs palmitoylation unresolved at this stage\", \"Calcineurin inhibition mechanism only partially mapped\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Demonstrated that the scaffold assembles a true ternary kinase-scaffold-phosphatase complex and uses it to bidirectionally regulate AMPA receptors and L-type channels.\",\n      \"evidence\": \"Live-cell FRET (~5 nm), electrophysiology, NFAT reporter and phosphorylation assays in HEK293 and hippocampal neurons (multiple studies)\",\n      \"pmids\": [\"12507994\", \"11943807\", \"12114507\", \"12354762\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and oligomeric state of the complex not yet defined\", \"Trafficking effects on CaV1.2 PKA-independent — mechanism unclear at this point\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Connected the scaffold to excitation-transcription coupling, showing anchored calcineurin reads local L-type channel Ca2+ to activate NFAT.\",\n      \"evidence\": \"Co-IP, electrophysiology and NFAT reporter assays with dominant-negative AKAP in HEK293 and neurons\",\n      \"pmids\": [\"17640527\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How NFAT itself is recruited to the complex not yet defined\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Identified AKAP5 as the convergent platform for inflammatory TRPV1 sensitization, extending its role to nociception.\",\n      \"evidence\": \"Co-IP, TRPV1 C-terminus deletion mapping, electrophysiology and in vivo hyperalgesia assays\",\n      \"pmids\": [\"18701070\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative contribution of each anchored enzyme to sensitization not dissected here\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Broadened the scaffold's enzyme repertoire to adenylyl cyclases and extended its role to cardiac and GPCR signalosomes, establishing tissue-wide compartmentalized cAMP control.\",\n      \"evidence\": \"Co-IP, FRET, AC activity assays in AKAP150 KO brain, cardiac KO mice, and cAMP biosensors (multiple studies)\",\n      \"pmids\": [\"20231277\", \"20671242\", \"20410303\", \"20188672\", \"20428246\", \"20664520\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How AC isoform selectivity is achieved across tissues not fully mapped\", \"D36 vs full-KO phenotype divergence mechanism partly unexplained\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined the quaternary architecture and added PP1 regulation and palmitoylation-dependent raft targeting, explaining how the scaffold physically integrates multiple enzymes and is membrane-positioned.\",\n      \"evidence\": \"Native MS and cross-linking of reconstituted complex, SPR/phosphatase assays, palmitoylation mutagenesis with raft fractionation and AC8 assays (multiple studies)\",\n      \"pmids\": [\"21464287\", \"21561082\", \"21771783\", \"21273417\", \"21562284\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance of PP1 inhibition not established\", \"Which channel/receptor complexes depend on raft localization not fully cataloged\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Revealed a paradoxical affinity-tuning principle: the calcineurin-anchoring motif competes with NFAT's docking peptide, so AKAP5 affinity sets NFAT output, and PKA feedback phosphorylates AC8 to shape cAMP dynamics.\",\n      \"evidence\": \"Structural binding analysis with mutagenesis, NFAT/calcineurin recruitment assays, AC8 S112A mutagenesis with FRET cAMP imaging (multiple studies)\",\n      \"pmids\": [\"22343722\", \"22976297\", \"23259949\", \"22677788\", \"22693956\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether decoy-site tuning operates at endogenous expression levels unresolved\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Established AC anchoring (not just PKA anchoring) as the critical node for β-adrenergic GluA1 phosphorylation and LTP, and validated the TRPV1-binding motif as an analgesic target in vivo.\",\n      \"evidence\": \"AKAP5 KO and D36 knock-in mice with LTP electrophysiology; TAT-peptide hyperalgesia assays (multiple studies)\",\n      \"pmids\": [\"23649627\", \"23699529\", \"23889134\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which AC anchoring outweighs PKA anchoring not fully explained\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Showed AKAP5 governs homeostatic synaptic scaling and cardiac CaN/CaMKII balance, broadening its plasticity and disease roles.\",\n      \"evidence\": \"AKAP5 RNAi and GluA1 S845 knockin, trafficking/electrophysiology; AKAP5 KO mice with echocardiography (multiple studies)\",\n      \"pmids\": [\"25451194\", \"25225170\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Causal chain from AKAP5 loss to cardiac dilatation only partly resolved\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified DHHC2 as the enzyme controlling AKAP5 palmitoylation/endosomal targeting and showed scaffold coupling to STIM1/store-operated channels, linking lipid modification to plasticity and Ca2+ entry.\",\n      \"evidence\": \"DHHC2 RNAi with lipidated rescue mutant, spine imaging, electrophysiology; STIM1 T389 mutagenesis with ARC/CRAC recordings (multiple studies)\",\n      \"pmids\": [\"25589740\", \"25504574\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Regulation of DHHC2 activity itself unknown\", \"How STIM1 phosphorylation discriminates ARC vs CRAC at structural level not defined\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Defined the molecular switch for activity-dependent scaffold removal, showing CaMKII-driven depalmitoylation (not phosphorylation alone) drives synaptic removal during LTD, and that intrinsic disorder tunes anchored phosphatase drug sensitivity.\",\n      \"evidence\": \"CaMKII pharmacology and site mutagenesis with live spine imaging; negative-stain EM with live-cell sensors (multiple studies)\",\n      \"pmids\": [\"29196604\", \"28967377\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the depalmitoylating enzyme not established\", \"How conformational ensembles map to specific functional states unresolved\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Pinpointed the C-terminal leucine-zipper as the direct NFAT-recruitment determinant and assigned PKC-dependent GluA1 Ser831 phosphorylation to generation of Ca2+-permeable AMPARs.\",\n      \"evidence\": \"AKAP150 RNAi with LZ-mutant rescue, FRET/NFAT/Ca2+ assays; GluA1 phosphosite mutagenesis with I-V electrophysiology (multiple studies)\",\n      \"pmids\": [\"31091162\", \"30737285\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of LZ-NFAT contact not solved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Quantified how scaffolding accelerates calcineurin-mediated suppression of PKA and dissected the Orai1-AKAP5-NFAT cytokine pathway, giving a kinetic and structural account of excitation-transcription coupling and PKA antagonism.\",\n      \"evidence\": \"In vitro reconstitution phosphatase assays with PKA reporters and LTD electrophysiology; NMR of the Orai1 N-terminus; cAMP nexus mapping; β-cell leptin FRET reporters (multiple studies)\",\n      \"pmids\": [\"34612814\", \"33941685\", \"34458850\", \"33617875\", \"33082339\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether the ~10-fold calcineurin acceleration occurs at endogenous stoichiometry in vivo not confirmed\", \"AC requirement for Orai1-linked cAMP appears cell-type-dependent\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Established dynamic exchange kinetics of AKAP5 on Orai1, explaining how transient scaffold occupancy and NFAT recycling sustain transcription during continuous Ca2+ entry.\",\n      \"evidence\": \"FRAP, FCS, co-IP and NFAT1 translocation with mathematical kinetic modeling\",\n      \"pmids\": [\"36317924\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab kinetic model awaits independent confirmation\", \"Molecular driver of the rapid AKAP5 exchange not identified\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how the many enzyme and channel modules are dynamically prioritized at a single AKAP5 scaffold within a cell, and which depalmitoylase and upstream signals control the activity-dependent assembly/disassembly cycle in vivo.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No full-length high-resolution structure of an assembled signalosome on a native channel\", \"Depalmitoylating enzyme unidentified\", \"Selectivity rules governing which partner complex forms in a given compartment are unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 11, 25, 44]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [5, 10, 27, 30, 48]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [3, 28]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [3, 42]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3, 11, 20, 46]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [40]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [1, 11, 42]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [23, 53]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 6, 13, 14, 37, 52]},\n      {\"term_id\": \"R-HSA-112316\", \"supporting_discovery_ids\": [8, 17, 36, 38, 42]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [16, 31, 44, 49]},\n      {\"term_id\": \"R-HSA-397014\", \"supporting_discovery_ids\": [20, 39, 46]}\n    ],\n    \"complexes\": [\n      \"AKAP79-PKA(RII)-calcineurin(PP2B)-CaM quaternary complex\",\n      \"caveolin-3/AC5-6/PKA/calcineurin/CaV1.2 cardiac T-tubule complex\",\n      \"RXFP1-AKAP79-AC2-β-arrestin2-PDE4D3 signalosome\",\n      \"Orai1-AKAP79-calcineurin-NFAT1 complex\"\n    ],\n    \"partners\": [\n      \"PRKAR2 (PKA RII)\",\n      \"PPP3CA (calcineurin A)\",\n      \"PRKCB (PKC)\",\n      \"CALM1 (calmodulin)\",\n      \"ADCY8 (AC8)\",\n      \"DLG1 (SAP97)\",\n      \"CACNA1C (CaV1.2)\",\n      \"ORAI1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}