{"gene":"ZDHHC5","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":2010,"finding":"DHHC5 co-immunoprecipitates with PSD-95 in neurons; this interaction is mediated through binding of the C-terminus of DHHC5 to the PDZ3 domain of PSD-95. DHHC5 is highly enriched in post-synaptic density preparations and localizes to CA3 and dentate gyrus of the hippocampus.","method":"Co-immunoprecipitation, subcellular fractionation (PSD preparation), immunohistochemistry","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP with domain mapping, single lab, multiple methods","pmids":["20178993"],"is_preprint":false},{"year":2011,"finding":"DHHC5 palmitoylates flotillin-2; palmitoylation and oligomerization of flotillin-2 are abolished in DHHC5-deficient neuronal stem cells, and overexpression of DHHC5 in COS-1 cells markedly stimulates flotillin-2 palmitoylation, establishing a direct enzyme-substrate relationship.","method":"Bioorthogonal palmitate analog (17-octadecynoic acid) metabolic labeling, quantitative proteomics, overexpression in COS-1 cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (metabolic labeling + overexpression rescue), single lab","pmids":["22081607"],"is_preprint":false},{"year":2011,"finding":"DHHC5 down-regulation is triggered within minutes of growth factor withdrawal from neural stem cells (a signal for neuronal differentiation), and this degradation is partially prevented by inhibitors of ubiquitin-mediated proteolysis, indicating DHHC5 protein levels are regulated by ubiquitin-dependent degradation in response to differentiation signals.","method":"Growth factor withdrawal experiments, proteasome inhibitor treatment, time-course protein level analysis","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct pharmacological intervention with defined readout, single lab","pmids":["22081607"],"is_preprint":false},{"year":2011,"finding":"ZDHHC5 palmitoylates somatostatin receptor 5 (SSTR5); ZDHHC5 interacts with the membrane-proximal helix 8 of SSTR5 and co-localizes with it at the plasma membrane. Overexpression of ZDHHC5 increases SSTR5 palmitoylation and knockdown decreases it, identifying ZDHHC5 as the first palmitoyltransferase for a GPCR.","method":"Ras recruitment interaction screen, co-immunoprecipitation, siRNA knockdown, palmitoylation assay","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus gain- and loss-of-function palmitoylation assays, single lab","pmids":["21820437"],"is_preprint":false},{"year":2012,"finding":"DHHC5 (together with DHHC8) palmitoylates GRIP1b via a PDZ ligand unique to DHHC5/8. Palmitoylated GRIP1b is targeted to trafficking endosomes, where it links endosomes to kinesin motors and accelerates AMPA receptor recycling. GRIP1b palmitoylation turnover rate is among the highest reported.","method":"Binding assays, palmitoylation assays, live imaging of dendritic endosomes, AMPA-R recycling assays","journal":"Neuron","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (binding, palmitoylation, trafficking, functional receptor assay), replicated with domain mutants","pmids":["22325201"],"is_preprint":false},{"year":2014,"finding":"DHHC5 palmitoylates phospholemman (PLM) at cysteines C40 and C42 (C40 being the principal site) in cardiac ventricular myocytes. DHHC5 localizes to caveolin-enriched cell surface microdomains and co-immunoprecipitates with PLM. Substrate recruitment requires a ~120 amino acid region of the DHHC5 C-tail immediately after the fourth transmembrane domain, not its PDZ-binding motif. PLM palmitoylation at C40, but not C42, is required for PLM-mediated inhibition of the Na+/K+ ATPase.","method":"Co-immunoprecipitation, overexpression and siRNA silencing, site-directed mutagenesis (C40A, C42A), Na pump activity assay, subcellular fractionation","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — mutagenesis of both enzyme (C-tail deletion) and substrate (C40A/C42A) combined with functional Na pump assay, multiple orthogonal methods","pmids":["25422474"],"is_preprint":false},{"year":2014,"finding":"DHHC5 transiently palmitoylates δ-catenin after enhanced synaptic activity, promoting δ-catenin-cadherin interactions at synapses. This palmitoylation is required for activity-induced stabilization of N-cadherin at synapses, enlargement of postsynaptic spines, insertion of GluA1/GluA2 into the synaptic membrane, and increased mEPSC amplitude. Context-dependent fear conditioning in mice increased δ-catenin palmitoylation and δ-catenin-cadherin associations at hippocampal synapses.","method":"Palmitoylation assay, co-immunoprecipitation, spine morphology imaging, electrophysiology (mEPSC), in vivo fear conditioning","journal":"Nature neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (biochemistry, imaging, electrophysiology, in vivo), replicated across cell and animal contexts","pmids":["24562000"],"is_preprint":false},{"year":2015,"finding":"Under basal conditions, DHHC5 is bound to PSD-95 and Fyn kinase at the synaptic membrane; Fyn-mediated phosphorylation of a tyrosine residue within the endocytic motif of DHHC5 stabilizes it at the membrane. Neuronal activity disrupts the DHHC5/PSD-95/Fyn complex, causing DHHC5 endocytosis and translocation to dendritic shafts where it encounters and palmitoylates its substrate δ-catenin, after which both return to spines.","method":"Live-cell imaging, co-immunoprecipitation, pharmacological inhibition of Fyn, endocytosis assays, palmitoylation assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (live imaging, Co-IP, pharmacological perturbation, palmitoylation), mechanistic model confirmed by functional readouts","pmids":["26334723"],"is_preprint":false},{"year":2015,"finding":"Catalytic activity of DHHC5 is required for NSCLC cell proliferation, colony formation, invasion, and tumor xenograft growth; rescue with wild-type but not catalytically inactive DHHC5 confirms the dependence on enzymatic activity.","method":"siRNA/shRNA knockdown, lentiviral re-expression of WT vs. catalytic mutant DHHC5, in vitro proliferation/invasion assays, xenograft mouse models","journal":"Molecular cancer research : MCR","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — catalytic mutant rescue experiment in both cell culture and in vivo, single lab","pmids":["25573953"],"is_preprint":false},{"year":2017,"finding":"ZDHHC5 palmitoylates EZH2 in glioma cells; mutant p53 transcriptionally upregulates ZDHHC5 (together with NF-Y), which alters EZH2 palmitoylation and phosphorylation status, promoting glioma stem-like cell self-renewal and tumorigenicity.","method":"Co-immunoprecipitation, palmitoylation assay, ChIP/reporter assay for transcriptional regulation, glioma stem cell self-renewal assays","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple methods (palmitoylation assay, transcriptional mechanism, functional stem cell assay), single lab","pmids":["28775165"],"is_preprint":false},{"year":2017,"finding":"DHHC5 palmitoylates the S1P receptor subtype 1 (S1P1R) at the plasma membrane, which is prerequisite for Gi protein coupling. Upon receptor stimulation and internalization, S1P1R is depalmitoylated. FTY720-P-induced endocytosed S1P1R vesicles subsequently associate with DHHC5, enabling persistent intracellular Gi signaling.","method":"Palmitoylation assay, co-immunoprecipitation, subcellular localization imaging, cAMP/Gi signaling assays, receptor internalization assays","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — palmitoylation assay linked to functional G-protein coupling readout, multiple methods, single lab","pmids":["29185452"],"is_preprint":false},{"year":2019,"finding":"DHHC4 and DHHC5 palmitoylate CD36 at different subcellular localizations to control its plasma membrane localization and fatty acid uptake activity. Depletion of DHHC5 disrupts CD36-dependent fatty acid uptake in cells, and adipose-specific Dhhc5 knockout mice show decreased fatty acid uptake in adipose tissue and develop hypothermia upon cold exposure.","method":"Palmitoylation assay, siRNA/shRNA knockdown, fatty acid uptake assay, conditional knockout mouse model, cold exposure phenotype","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — cell-based and in vivo knockout with multiple mechanistic readouts (palmitoylation, localization, function), replicated with two enzymes","pmids":["30605677"],"is_preprint":false},{"year":2019,"finding":"ZDHHC5 forms a catalytically active protein S-acyltransferase complex with the accessory subunit GOLGA7. The ZDHHC5-GOLGA7 complex is mutually stabilizing and localizes to the plasma membrane. This complex is required for CIL56-induced nonapoptotic cell death and is involved in retrograde protein trafficking from the plasma membrane.","method":"CRISPR genetic screen, co-immunoprecipitation, subcellular localization imaging, cell death assays with catalytic mutants","journal":"Cell chemical biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR screen plus Co-IP and catalytic mutant validation, single lab","pmids":["31631010"],"is_preprint":false},{"year":2019,"finding":"DHHC5 palmitoylates the proprotein convertases Furin and PC7. ZDHHC5-mediated palmitoylation promotes Furin/PC7 association with plasma membrane microdomains, which is required for cleavage of anthrax toxin protective antigen (but does not inhibit protease activity per se). ZDHHC5 also affects homeostasis and trafficking of Furin/PC7 through the endocytic/recycling pathway.","method":"Palmitoylation assay, subcellular fractionation (lipid raft), siRNA knockdown, anthrax toxin processing assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — palmitoylation assay plus functional toxin processing readout, single lab, multiple methods","pmids":["30610172"],"is_preprint":false},{"year":2019,"finding":"DHHC5 binds to and palmitoylates Golga7b. Palmitoylation of Golga7b prevents clathrin-mediated endocytosis of DHHC5 and stabilizes it at the plasma membrane. Desmoglein-2 and plakophilin-3 are substrates of DHHC5, and DHHC5/Golga7b are required for desmoglein-2 plasma membrane localization, desmosomal patterning, and cell adhesion.","method":"Co-immunoprecipitation, palmitoylation assay, siRNA knockdown, fluorescence imaging, proteomic analysis of complexes, cell adhesion assays","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (Co-IP, palmitoylation assay, proteomics, imaging, functional adhesion assay), multiple substrates identified","pmids":["31402609"],"is_preprint":false},{"year":2019,"finding":"DHHC5 mediates β-adrenergic stimulation-dependent palmitoylation of Gαs and Gαi in cardiomyocytes. An in vitro palmitoylation assay with purified components established Gαs and Gαi as direct DHHC5 substrates. The C-terminal tail of DHHC5 itself is palmitoylated in response to stimulation, which is important for DHHC5 dynamic localization and function at the plasma membrane.","method":"In vitro palmitoylation assay with purified components, siRNA knockdown, cAMP measurement, contractility assay, subcellular localization imaging","journal":"Biophysical journal","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with purified components plus functional cellular readouts, single lab","pmids":["31547976"],"is_preprint":false},{"year":2020,"finding":"Among all mammalian PATs, only ZDHHC5 and ZDHHC8 are enriched in DRG axons. Both are required for Gp130/JAK/STAT3 (but not DLK/JNK) axon-to-soma retrograde signaling after injury. ZDHHC5 and ZDHHC8 palmitoylate Gp130 in co-transfected cells; shRNA knockdown reduces Gp130 palmitoylation and surface expression in DRG neurons.","method":"Immunofluorescence localization screen of all PATs in DRG neurons, shRNA knockdown, palmitoylation assay, STAT3/JNK signaling assays, co-transfection palmitoylation assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — comprehensive PAT localization screen plus palmitoylation and signaling assays, single lab","pmids":["32958558"],"is_preprint":false},{"year":2021,"finding":"DHHC5 palmitoylates STAT3 in oligodendrocytes; STAT3 is the primary DHHC5 substrate in these cells. DHHC5 ablation reduces STAT3 palmitoylation and suppresses STAT3 phosphorylation/activation, inhibiting transcription of myelin-related and anti-apoptosis genes and suppressing myelination and remyelination.","method":"Conditional Zdhhc5 knockout, palmitoylation assay, STAT3 phosphorylation western blot, myelination histology, gene expression analysis","journal":"Glia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — conditional KO with mechanistic substrate identification and functional myelination readout, single lab","pmids":["34724258"],"is_preprint":false},{"year":2021,"finding":"ZDHHC5 and GOLGA7 interact with the SARS-CoV-2 spike protein and are required for efficient SARS-CoV-2 pseudovirus entry into A549 and HeLa cells. ZDHHC5/GOLGA7 knockout had no significant effect on S protein subcellular localization or palmitoylation, but significantly decreased pseudovirus entry.","method":"CRISPR-Cas9 knockout, co-immunoprecipitation, ABE palmitoylation assay, pseudovirus entry luciferase assay, fluorescence microscopy","journal":"Virology journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR KO plus multiple assays, single lab; mechanism of entry effect not fully resolved","pmids":["34961524"],"is_preprint":false},{"year":2022,"finding":"DHHC5 palmitoylates the brain-specific isoform Cdc42-palm; DHHC5 physically interacts with Cdc42-palm. A dominant-negative DHHC5 mutant or DHHC5 knockdown decreases Cdc42-palm palmitoylation and plasma membrane localization, leading to reduced Cdc42-mediated gene transcription and spine formation in hippocampal neurons.","method":"Co-immunoprecipitation, palmitoylation assay, dominant-negative and shRNA approaches, fluorescence imaging, spine morphology analysis, transcription reporter assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus loss-of-function with multiple cellular readouts, single lab","pmids":["35597282"],"is_preprint":false},{"year":2023,"finding":"ZDHHC5 palmitoylates NLRP3 at the LRR domain. This palmitoylation promotes NLRP3 oligomerization, NLRP3-NEK7 interaction, and ASC aggregate formation, leading to caspase-1 activation, IL-1β/18 release, and GSDMD cleavage. ABHD17A acts as the depalmitoylase for NLRP3. Zdhhc5-/- mice show defective NLRP3 inflammasome activation in vivo.","method":"Palmitoylation assay, siRNA/KO, co-immunoprecipitation (NLRP3-NEK7), ASC speck imaging, caspase-1 activation assay, IL-1β/18 ELISA, GSDMD cleavage, Zdhhc5 KO mouse model","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (palmitoylation assay, protein-protein interaction, inflammasome assembly, in vivo KO), replicated in human cells and mice","pmids":["38092000"],"is_preprint":false},{"year":2023,"finding":"ZDHHC5 palmitoylates FAK at C456, promoting FAK membrane localization and phosphorylation (activation). ZDHHC5 knockdown abolishes FAK palmitoylation and membrane distribution, reducing cell proliferation, invasion, and EMT in glioblastoma cells and tumor growth in intracranial xenografts.","method":"Acyl-PEG exchange assay, metabolic incorporation assay, subcellular fractionation/imaging, siRNA knockdown, 2-BP inhibition, CCK8/invasion/colony assays, intracranial xenograft","journal":"Cell communication and signaling : CCS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — palmitoylation site identified by metabolic assay plus functional in vitro and in vivo readouts, single lab","pmids":["38233791"],"is_preprint":false},{"year":2023,"finding":"ZDHHC5 palmitoylates protocadherin 7 (PCDH7) in a cell cycle-dependent manner. PCDH7 and ZDHHC5 colocalize at the mitotic cell surface and translocate together to the cleavage furrow. PCDH7 localization at the cell surface depends on ZDHHC5 palmitoylation activity. Loss of PCDH7 increases multinucleated cells, prolongs mitosis, and reduces active RhoA and phospho-myosin at the cleavage furrow.","method":"Co-immunoprecipitation, palmitoylation assay, live-cell imaging, siRNA knockdown, RhoA activity assay, phospho-myosin immunofluorescence","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus palmitoylation-dependent localization and functional cytokinesis readouts, single lab","pmids":["36762613"],"is_preprint":false},{"year":2023,"finding":"ZDHHC5 palmitoylates SSTR5 and lomitapide, an FDA-approved drug, acts as an inhibitor of ZDHHC5, blocking palmitoylation of SSTR5 and reducing pancreatic cancer cell growth in vitro and in vivo.","method":"Palmitoylation assay, drug treatment (lomitapide), cell proliferation assays, in vivo xenograft","journal":"Cell death discovery","confidence":"Low","confidence_rationale":"Tier 3 / Weak — pharmacological inhibitor study with functional readout; mechanism of ZDHHC5 inhibition by lomitapide not fully characterized, single lab","pmids":["36774350"],"is_preprint":false},{"year":2023,"finding":"Frizzled-5 (Fz5) is S-acylated on three C-terminal cysteines by zDHHC5. S-acylation is essential for Fz5 localization to the cell surface, axons, and presynaptic sites; S-acylation-deficient Fz5 is internalized faster and fails to activate canonical and divergent canonical Wnt pathways. S-acylation levels of Fz5 are regulated by neuronal activity, and S-acylation-deficient Fz5 fails to induce presynaptic assembly in vivo.","method":"Palmitoylation assay, mutagenesis of acylation sites, live imaging, internalization assay, Wnt pathway reporter assay, in vivo presynaptic assembly assay","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 1 / Moderate — mutagenesis of acylation sites combined with functional pathway and in vivo synaptogenesis assays, multiple orthogonal methods","pmids":["37557176"],"is_preprint":false},{"year":2024,"finding":"DHHC5 mediates palmitoylation of beclin 1, which promotes formation of the ATG14L-containing class III PI3K complex I (PI3KC3-C1) and its lipid kinase activity, by enhancing hydrophobic interactions between beclin 1 and adapter proteins ATG14L and VPS15. DHHC5 deficiency in neurons exaggerates neurodegeneration in Alzheimer's disease mouse models in an autophagy-dependent manner.","method":"Palmitoylation assay, co-immunoprecipitation, PI3K lipid kinase activity assay, Zdhhc5 KO neuronal models, autophagy flux analysis, AD mouse models","journal":"Nature structural & molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — enzymatic activity assay (PI3K) linked directly to palmitoylation, multiple orthogonal methods, disease-relevant in vivo models","pmids":["38177673"],"is_preprint":false},{"year":2024,"finding":"ZDHHC5 palmitoylates PKCδ in hypothalamic microglia; an antimalarial drug (artemether) blocks ZDHHC5-PKCδ binding to inhibit PKCδ palmitoylation, suppressing downstream neuroinflammation signaling and improving hepatic lipid metabolism via TRH/thyroid hormone neuroendocrine axis.","method":"Biotin-labeled chemical probe pulldown, co-immunoprecipitation, palmitoylation assay, siRNA knockdown, neuroendocrine assays, high-fat diet mouse model","journal":"Theranostics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — chemical probe target ID plus Co-IP, palmitoylation assay, and in vivo KD, single lab","pmids":["38250049"],"is_preprint":false},{"year":2024,"finding":"ZDHHC5 palmitoylates SMPDL3B, and high glucose exposure increases ZDHHC5-SMPDL3B interaction. Palmitoylation by ZDHHC5 stabilizes SMPDL3B protein; inhibiting palmitoylation accelerates SMPDL3B degradation, while inhibiting depalmitoylation decreases its turnover rate in retinal endothelial cells.","method":"Co-immunoprecipitation, palmitoylation inhibitor (2-BP) and depalmitoylation inhibitor (palmostatin B) treatment, protein stability assay","journal":"Cellular signalling","confidence":"Low","confidence_rationale":"Tier 3 / Weak — co-IP and pharmacological inhibitors without direct palmitoylation site identification or reconstitution, single lab","pmids":["38266744"],"is_preprint":false},{"year":2024,"finding":"ZDHHC5 palmitoylates BRAF at Cys194/195 in cholangiocarcinoma cells, promoting BRAF membrane localization and protein stabilization. This activates ERK/MAPK signaling. Knockdown of ZDHHC5 or mutation of BRAF palmitoylation sites inhibits ERK signaling and cholangiocarcinoma growth.","method":"Palmitoylation assay, site-directed mutagenesis (C194/195A), ERK western blot, siRNA knockdown, cell proliferation assay","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — palmitoylation site mutagenesis plus functional signaling and growth assays, single lab","pmids":["42134490"],"is_preprint":false},{"year":2025,"finding":"DHHC5 palmitoylates transferrin receptor 1 (TfR1) at C98 in oligodendrocytes, which is required for regulation of TfR1 endocytosis. In neonatal sevoflurane-exposed mice, DHHC5 is specifically downregulated in oligodendrocytes, reducing TfR1 palmitoylation, enhancing TfR1 endocytosis, causing iron accumulation and ferroptosis, leading to hypomyelination.","method":"Acyl-resin assisted capture assay, co-immunoprecipitation, specific oligodendrocyte DHHC5 overexpression (Pdgfrα-CreERT), single-cell RNA sequencing, TfR1 endocytosis assay","journal":"Journal of advanced research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — palmitoylation site identified plus conditional rescue in specific cell type in vivo, single lab","pmids":["39929269"],"is_preprint":false},{"year":2025,"finding":"ZDHHC5 mediates IFT81 palmitoylation in round spermatids; ZDHHC5 predominantly localizes to the Golgi apparatus of round spermatids (steps 2-8). Zdhhc5 KO male mice are completely infertile with defective spermatogenesis, reduced sperm motility, and sperm tail malformations linked to defective IFT81 regulation.","method":"CRISPR/Cas9 knockout, immunofluorescence localization, palmitoylation assay, sperm morphology/motility analysis, IVF, ICSI rescue","journal":"Reproduction (Cambridge, England)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO with specific cellular localization and substrate identification plus functional rescue experiment (ICSI), single lab","pmids":["41182177"],"is_preprint":false},{"year":2025,"finding":"DHHC5 palmitoylates CRYBG1 (an actin-binding protein) in lymphatic endothelial cells; CRYBG1 palmitoylation is required for VEGFR2 lipid raft localization, which promotes VEGFR2 signaling and intestinal lymphatic integrity. Dhhc5 intestinal/LEC-specific knockout mice show impaired intestinal lipid absorption and resistance to diet-induced obesity.","method":"Conditional Dhhc5 knockout, palmitoylation assay, lipid raft fractionation, VEGFR2 signaling assay, lipid absorption assay","journal":"Life metabolism","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — conditional KO with mechanistic substrate identification and functional lipid absorption readout, single lab","pmids":["40589731"],"is_preprint":false},{"year":2025,"finding":"AMPK phosphorylates ZDHHC5 at Ser296 and Ser380, which weakens ZDHHC5 association with GOLGA7B and promotes ZDHHC5 displacement from the plasma membrane, thereby reducing NOD1 palmitoylation, NOD1 membrane localization, and downstream NOD1 innate immune signaling. Conversely, NOD1 agonist C12-iE-DAP suppresses AMPK activity to stabilize ZDHHC5 at the membrane.","method":"Biochemical phosphorylation assay, site-directed mutagenesis (Ser296/380), co-immunoprecipitation, palmitoylation assay, NOD1 membrane fractionation, NF-κB signaling assay","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — phosphorylation site mutagenesis plus functional NOD1 signaling readout, multiple methods, single lab","pmids":["41890956"],"is_preprint":false},{"year":2025,"finding":"ZDHHC5 palmitoylates NADK at Cys22, Cys23, and Cys26 within its amino-terminal domain, stimulating NADK kinase activity by relieving an autoinhibitory function of the amino terminus and promoting NADP+ synthesis. Zdhhc5-/- mice show defective NADK palmitoylation and NADP+ production.","method":"In vitro palmitoylation assay, site-directed mutagenesis, NADK kinase activity assay, Zdhhc5 KO mouse model, metabolomics (NADP+ measurement)","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — in vitro assay with mutagenesis and in vivo KO, but preprint (not peer-reviewed), single lab","pmids":[],"is_preprint":true},{"year":2025,"finding":"The ZDHHC5-GOLGA7 complex structure was determined by cryo-EM. Key conserved residues in both ZDHHC5 and GOLGA7 required for complex formation were identified by mutagenesis; disruption of these residues abolished CIL56-induced nonapoptotic cell death.","method":"Cryo-EM structure determination, biochemical purification, homology modeling, mutagenesis, cell death functional assays","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — cryo-EM structure with mutagenesis validation, but preprint (not peer-reviewed), single lab","pmids":[],"is_preprint":true},{"year":2025,"finding":"DHHC5 palmitoylates Syk on a single cysteine residue during FcγR-mediated phagocytosis in macrophages. Syk palmitoylation is required for Syk localization to the phagocytic cup, Syk phosphorylation/activation, Cdc42 recruitment, F-actin polymerization, and phagocytosis.","method":"S-acylation assay, site-directed mutagenesis, fluorescence imaging of phagocytic cup, Syk phosphorylation western blot, phagocytosis assay","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — multiple mechanistic readouts with mutagenesis, but preprint (not peer-reviewed), single lab","pmids":[],"is_preprint":true},{"year":2025,"finding":"ZDHHC5 palmitoylates YBX1 (via PANoptosis-related mechanism in intervertebral disc cells), and ZDHHC5-enriched extracellular vesicles modulate ZBP1 transcription through competitive inhibition of YBX1 phosphorylation via palmitoylation.","method":"Transcriptomics, scRNA-seq, co-immunoprecipitation, palmitoylation assay, extracellular vesicle delivery, transcriptional reporter assay","journal":"Journal of tissue engineering","confidence":"Low","confidence_rationale":"Tier 3 / Weak — mechanism proposed from EV-delivery experiments; palmitoylation of YBX1 by ZDHHC5 not directly confirmed by reconstitution, single lab","pmids":["41179831"],"is_preprint":false}],"current_model":"ZDHHC5 is a plasma membrane-localized DHHC-family palmitoyl acyltransferase that S-palmitoylates a broad range of substrates—including CD36, GRIP1b, δ-catenin, PLM, flotillin-2, NLRP3, Gαs/Gαi, beclin 1, EZH2, STAT3, FAK, BRAF, S1P1R, Fz5, SSTR5, NOD1, Cdc42, Syk, PCDH7, Furin/PC7, Gp130, desmoglein-2, IFT81, CRYBG1, TfR1, and NADK—thereby controlling their membrane localization, stability, activity, and interactions; ZDHHC5 activity is itself regulated by Fyn-mediated tyrosine phosphorylation of its endocytic motif (controlling activity-dependent neuronal trafficking), AMPK-mediated phosphorylation at Ser296/380 (coupling metabolic state to innate immune signaling), palmitoylation of its own C-terminal tail, and complex formation with accessory proteins GOLGA7/Golga7b that stabilize it at the plasma membrane; through these mechanisms ZDHHC5 governs diverse processes including synaptic plasticity, AMPA receptor recycling, cardiac sodium pump regulation, β-adrenergic signaling, NLRP3 inflammasome assembly, autophagy, myelination, fatty acid uptake, cytokinesis, axonal retrograde signaling, and cell adhesion."},"narrative":{"mechanistic_narrative":"ZDHHC5 is a plasma-membrane DHHC-family protein S-acyltransferase that palmitoylates a broad set of substrates to control their membrane localization, stability, and activity, thereby governing synaptic plasticity, innate immunity, metabolism, and cell adhesion [PMID:22325201, PMID:25422474, PMID:38092000]. At neuronal synapses it is held at the membrane through a complex with PSD-95 and Fyn, in which Fyn-mediated tyrosine phosphorylation of the ZDHHC5 endocytic motif stabilizes it; neuronal activity disrupts this complex and drives ZDHHC5 endocytosis to dendritic shafts where it transiently palmitoylates δ-catenin to stabilize cadherin and remodel spines [PMID:24562000, PMID:26334723], and it likewise palmitoylates GRIP1b to accelerate AMPA-receptor recycling [PMID:22325201] and Cdc42-palm to support spine formation [PMID:35597282]. Its enzymatic activity is required for substrate effects, demonstrated by catalytically inactive rescue failures across multiple systems [PMID:25422474, PMID:25573953]. ZDHHC5 is stabilized at the plasma membrane by the accessory subunit GOLGA7, with which it forms a mutually stabilizing, catalytically active complex, and by palmitoylation of GOLGA7b that blocks its clathrin-mediated endocytosis [PMID:31631010, PMID:31402609]; its own C-terminal tail is also palmitoylated in a stimulus-dependent manner to tune membrane dynamics [PMID:31547976]. Through palmitoylation of NLRP3 it promotes inflammasome oligomerization, NLRP3-NEK7 interaction, and caspase-1 activation, with ABHD17A serving as the opposing depalmitoylase [PMID:38092000], and AMPK phosphorylation at Ser296/Ser380 weakens its GOLGA7B association to dampen NOD1 signaling, coupling metabolic state to innate immunity [PMID:41890956]. ZDHHC5 also palmitoylates beclin 1 to assemble the ATG14L-containing PI3KC3-C1 autophagy complex [PMID:38177673], CD36 to drive fatty acid uptake [PMID:30605677], desmoglein-2 to pattern desmosomes and mediate adhesion [PMID:31402609], and signaling effectors including BRAF, FAK, STAT3, and Gp130 that link it to MAPK/ERK, STAT3, and retrograde axonal signaling and to tumor and myelination phenotypes [PMID:32958558, PMID:34724258, PMID:38233791, PMID:42134490].","teleology":[{"year":2010,"claim":"Established where and with what ZDHHC5 associates in the brain, anchoring it physically to the postsynaptic density rather than treating it as a diffuse membrane enzyme.","evidence":"Reciprocal Co-IP with domain mapping, PSD fractionation, and immunohistochemistry in neurons","pmids":["20178993"],"confidence":"Medium","gaps":["Did not identify catalytic substrates","Functional consequence of the PSD-95 interaction unresolved at this stage"]},{"year":2011,"claim":"Identified the first concrete substrates (flotillin-2, SSTR5) and showed ZDHHC5 protein levels are themselves controlled by ubiquitin-dependent degradation, establishing it as a regulated enzyme with defined targets.","evidence":"Metabolic palmitate labeling, overexpression rescue, Ras-recruitment interaction screen, and proteasome inhibitor treatment in neural stem cells and COS-1 cells","pmids":["22081607","21820437"],"confidence":"Medium","gaps":["Palmitoylation site residues not mapped","Mechanism of differentiation-triggered degradation not defined"]},{"year":2012,"claim":"Connected ZDHHC5 catalysis to a defined neuronal trafficking output by showing GRIP1b palmitoylation routes endosomes to kinesin motors and accelerates AMPA-receptor recycling.","evidence":"Binding and palmitoylation assays, live imaging of dendritic endosomes, AMPA-R recycling assays","pmids":["22325201"],"confidence":"High","gaps":["Redundancy with DHHC8 not fully separated","Endogenous regulation of this turnover not addressed"]},{"year":2014,"claim":"Resolved how ZDHHC5 selects substrates and linked its activity to physiology, mapping a substrate-recruitment region in the C-tail and showing site-specific PLM palmitoylation controls Na+/K+ ATPase regulation, plus activity-dependent δ-catenin palmitoylation driving synaptic plasticity.","evidence":"Enzyme C-tail deletion and substrate C40A/C42A mutagenesis with Na pump assay in cardiomyocytes; palmitoylation, spine imaging, mEPSC, and in vivo fear conditioning","pmids":["25422474","24562000"],"confidence":"High","gaps":["Structural basis of C-tail substrate recruitment unknown","How activity is transduced to palmitoylation not yet mechanistic"]},{"year":2015,"claim":"Defined the trafficking control logic of ZDHHC5 itself, showing a Fyn/PSD-95-phosphorylation switch governs its membrane retention versus endocytic relocation to reach substrates, and demonstrated strict dependence on catalytic activity for oncogenic phenotypes.","evidence":"Live imaging, Co-IP, Fyn pharmacological inhibition, and endocytosis assays; catalytic-mutant rescue in NSCLC cells and xenografts","pmids":["26334723","25573953"],"confidence":"High","gaps":["Identity of the cancer-relevant substrate(s) not defined in the NSCLC study","Generality of the Fyn switch to non-neuronal cells unknown"]},{"year":2017,"claim":"Extended ZDHHC5 into GPCR signaling persistence and tumor biology, showing S1P1R palmitoylation enables Gi coupling and that mutant p53 transcriptionally upregulates ZDHHC5 to alter EZH2 in glioma stem cells.","evidence":"Palmitoylation assays with Gi signaling and internalization readouts; ChIP/reporter assays and stem-cell self-renewal assays","pmids":["29185452","28775165"],"confidence":"Medium","gaps":["EZH2 palmitoylation sites not mapped","Direct enzyme-substrate proof for EZH2 limited to Co-IP/palmitoylation"]},{"year":2019,"claim":"Established the GOLGA7/Golga7b accessory module that stabilizes ZDHHC5 at the membrane and broadly expanded its substrate range to metabolism (CD36), adhesion (desmoglein-2), secretion (Furin/PC7), and G-protein/self-palmitoylation, including in vivo knockout phenotypes.","evidence":"CRISPR screen, Co-IP, conditional Dhhc5 KO mice with cold-exposure and fatty-acid-uptake phenotypes, in vitro palmitoylation with purified Gα components, adhesion and toxin-processing assays","pmids":["31631010","30605677","31402609","30610172","31547976"],"confidence":"High","gaps":["Structural basis of the ZDHHC5-GOLGA7 complex not yet resolved at this stage","Rules governing which substrate is engaged in which compartment unclear"]},{"year":2021,"claim":"Demonstrated cell-type-specific dominant substrates, with STAT3 as the principal oligodendrocyte target controlling myelination through palmitoylation-dependent STAT3 activation.","evidence":"Conditional Zdhhc5 knockout, palmitoylation assay, STAT3 phosphorylation, and myelination histology","pmids":["34724258"],"confidence":"Medium","gaps":["STAT3 palmitoylation site not specified","How palmitoylation couples to STAT3 phosphorylation mechanistically unclear"]},{"year":2023,"claim":"Positioned ZDHHC5 as a central node in inflammasome assembly and cytokinesis and as a druggable cancer target, showing LRR-domain NLRP3 palmitoylation (opposed by ABHD17A) drives caspase-1 activation, FAK and BRAF palmitoylation drive tumor signaling, and PCDH7 palmitoylation supports cleavage-furrow function.","evidence":"Palmitoylation and site-directed mutagenesis, NLRP3-NEK7 Co-IP, ASC speck and caspase-1 assays, Zdhhc5 KO mice, RhoA/phospho-myosin and xenograft assays, lomitapide inhibitor study","pmids":["38092000","38233791","36762613","36774350"],"confidence":"High","gaps":["Lomitapide's direct mode of ZDHHC5 inhibition not characterized (Low-confidence)","Whether a single ZDHHC5 conformation handles such diverse substrates is unresolved"]},{"year":2024,"claim":"Linked ZDHHC5 to autophagy machinery assembly and additional metabolic/neuroinflammatory axes, showing beclin 1 palmitoylation builds the ATG14L PI3KC3-C1 complex with disease relevance in Alzheimer's models.","evidence":"Palmitoylation assay, PI3K lipid-kinase activity assay, Zdhhc5 KO neuronal and AD mouse models; PKCδ and SMPDL3B substrate studies","pmids":["38177673","38250049","38266744"],"confidence":"High","gaps":["SMPDL3B palmitoylation site and direct catalysis not established (Low-confidence)","Beclin 1 palmitoylation site within the corpus not detailed"]},{"year":2025,"claim":"Provided the first ZDHHC5-GOLGA7 cryo-EM structure and a metabolic regulatory loop (AMPK phosphorylation at Ser296/380 weakening GOLGA7B binding to dampen NOD1 signaling), plus new substrates spanning iron handling, lymphatic lipid uptake, NADK activation, spermatogenesis, and phagocytosis.","evidence":"Cryo-EM and mutagenesis (preprint), AMPK phosphosite mutagenesis with NOD1 signaling, conditional KO and KO mouse models, in vitro NADK kinase assay (preprint), Syk phagocytosis assay (preprint)","pmids":["41890956","39929269","40589731","41182177"],"confidence":"Medium","gaps":["Key structural and NADK/Syk findings remain in preprint form","A unifying model for compartment- and stimulus-specific substrate selection is still absent"]},{"year":null,"claim":"It remains unknown how a single ZDHHC5 enzyme achieves selectivity across its very large substrate set and how its localization, GOLGA7/GOLGA7b association, phosphorylation, and self-palmitoylation are integrated to direct it to the right substrate in the right compartment.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No comprehensive substrate-recognition code defined","Limited structural insight into catalysis-substrate engagement","Cross-talk between the multiple regulatory inputs not reconstituted"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[4,5,11,14,20,25]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[1,5,15,24]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[20,25,33]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[3,5,12,14,15]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[30]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[10,12]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[20,32]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[25]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[10,16,28]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[11,31]},{"term_id":"R-HSA-112316","term_label":"Neuronal System","supporting_discovery_ids":[4,6,7]}],"complexes":["ZDHHC5-GOLGA7 S-acyltransferase complex","DHHC5/PSD-95/Fyn synaptic complex"],"partners":["GOLGA7","GOLGA7B","PSD-95","FYN","NLRP3","CD36","NEK7","BECN1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9C0B5","full_name":"Palmitoyltransferase ZDHHC5","aliases":["Zinc finger DHHC domain-containing protein 5","DHHC-5","Zinc finger protein 375"],"length_aa":715,"mass_kda":77.5,"function":"Palmitoyltransferase that catalyzes the addition of palmitate onto various protein substrates such as CTNND2, CD36, GSDMD, NLRP3, NOD1, NOD2, STAT3 and S1PR1 thus plays a role in various biological processes including cell adhesion, inflammation, fatty acid uptake, bacterial sensing or cardiac functions (PubMed:21820437, PubMed:29185452, PubMed:31402609, PubMed:31649195, PubMed:34293401, PubMed:38092000, PubMed:38530158, PubMed:38599239). Plays an important role in the regulation of synapse efficacy by mediating palmitoylation of delta-catenin/CTNND2, thereby increasing synaptic delivery and surface stabilization of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors (AMPARs) (PubMed:26334723). Under basal conditions, remains at the synaptic membrane through FYN-mediated phosphorylation that prevents association with endocytic proteins (PubMed:26334723). Neuronal activity enhances the internalization and trafficking of DHHC5 from spines to dendritic shafts where it palmitoylates delta-catenin/CTNND2 (PubMed:26334723). Regulates cell adhesion at the plasma membrane by palmitoylating GOLGA7B and DSG2 (PubMed:31402609). Plays a role in innate immune response by mediating the palmitoylation of NOD1 and NOD2 and their proper recruitment to the bacterial entry site and phagosomes (PubMed:31649195, PubMed:34293401). Also participates in fatty acid uptake by palmitoylating CD36 and thereby targeting it to the plasma membrane (PubMed:32958780). Upon binding of fatty acids to CD36, gets phosphorylated by LYN leading to inactivation and subsequent CD36 caveolar endocytosis (PubMed:32958780). Controls oligodendrocyte development by catalyzing STAT3 palmitoylation (By similarity). Acts as a regulator of inflammatory response by mediating palmitoylation of NLRP3 and GSDMD (PubMed:38092000, PubMed:38530158, PubMed:38599239). Palmitoylates NLRP3 to promote inflammasome assembly and activation (PubMed:38092000). Activates pyroptosis by catalyzing palmitoylation of gasdermin-D (GSDMD), thereby promoting membrane translocation and pore formation of GSDMD (PubMed:38530158, PubMed:38599239)","subcellular_location":"Cell membrane; Synapse","url":"https://www.uniprot.org/uniprotkb/Q9C0B5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ZDHHC5","classification":"Not Classified","n_dependent_lines":12,"n_total_lines":1208,"dependency_fraction":0.009933774834437087},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"GOLGA7","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/ZDHHC5","total_profiled":1310},"omim":[{"mim_id":"617042","title":"GASDERMIN D; GSDMD","url":"https://www.omim.org/entry/617042"},{"mim_id":"614586","title":"ZDHHC PALMITOYLTRANSFERASE 5; ZDHHC5","url":"https://www.omim.org/entry/614586"},{"mim_id":"182455","title":"SOMATOSTATIN RECEPTOR 5; SSTR5","url":"https://www.omim.org/entry/182455"},{"mim_id":"131560","title":"FLOTILLIN 2; FLOT2","url":"https://www.omim.org/entry/131560"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Plasma membrane","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ZDHHC5"},"hgnc":{"alias_symbol":["KIAA1748","ZNF375","DHHC5"],"prev_symbol":[]},"alphafold":{"accession":"Q9C0B5","domains":[{"cath_id":"-","chopping":"10-244","consensus_level":"medium","plddt":92.6735,"start":10,"end":244}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9C0B5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9C0B5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9C0B5-F1-predicted_aligned_error_v6.png","plddt_mean":58.81},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ZDHHC5","jax_strain_url":"https://www.jax.org/strain/search?query=ZDHHC5"},"sequence":{"accession":"Q9C0B5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9C0B5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9C0B5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9C0B5"}},"corpus_meta":[{"pmid":"30605677","id":"PMC_30605677","title":"DHHC4 and DHHC5 Facilitate Fatty Acid Uptake by Palmitoylating and Targeting CD36 to the Plasma Membrane.","date":"2019","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/30605677","citation_count":162,"is_preprint":false},{"pmid":"22325201","id":"PMC_22325201","title":"Palmitoylation by DHHC5/8 targets GRIP1 to dendritic endosomes to regulate AMPA-R trafficking.","date":"2012","source":"Neuron","url":"https://pubmed.ncbi.nlm.nih.gov/22325201","citation_count":158,"is_preprint":false},{"pmid":"38092000","id":"PMC_38092000","title":"ZDHHC5-mediated NLRP3 palmitoylation promotes NLRP3-NEK7 interaction and inflammasome activation.","date":"2023","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/38092000","citation_count":122,"is_preprint":false},{"pmid":"22081607","id":"PMC_22081607","title":"DHHC5 protein palmitoylates flotillin-2 and is rapidly degraded on induction of neuronal differentiation in cultured cells.","date":"2011","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22081607","citation_count":122,"is_preprint":false},{"pmid":"28775165","id":"PMC_28775165","title":"EZH2 Palmitoylation Mediated by ZDHHC5 in p53-Mutant Glioma Drives Malignant Development and Progression.","date":"2017","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/28775165","citation_count":106,"is_preprint":false},{"pmid":"25422474","id":"PMC_25422474","title":"Substrate recognition by the cell surface palmitoyl transferase DHHC5.","date":"2014","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/25422474","citation_count":105,"is_preprint":false},{"pmid":"24562000","id":"PMC_24562000","title":"Palmitoylation of δ-catenin by DHHC5 mediates activity-induced synapse plasticity.","date":"2014","source":"Nature neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/24562000","citation_count":103,"is_preprint":false},{"pmid":"26334723","id":"PMC_26334723","title":"Activity-regulated trafficking of the palmitoyl-acyl transferase DHHC5.","date":"2015","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/26334723","citation_count":97,"is_preprint":false},{"pmid":"20178993","id":"PMC_20178993","title":"DHHC5 interacts with PDZ domain 3 of post-synaptic density-95 (PSD-95) protein and plays a role in learning and memory.","date":"2010","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20178993","citation_count":73,"is_preprint":false},{"pmid":"31631010","id":"PMC_31631010","title":"A ZDHHC5-GOLGA7 Protein Acyltransferase Complex Promotes Nonapoptotic Cell Death.","date":"2019","source":"Cell chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/31631010","citation_count":55,"is_preprint":false},{"pmid":"31402609","id":"PMC_31402609","title":"S-acylated Golga7b stabilises DHHC5 at the plasma membrane to regulate cell adhesion.","date":"2019","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/31402609","citation_count":55,"is_preprint":false},{"pmid":"25573953","id":"PMC_25573953","title":"Systematic siRNA Screen Unmasks NSCLC Growth Dependence by Palmitoyltransferase DHHC5.","date":"2015","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/25573953","citation_count":45,"is_preprint":false},{"pmid":"38177673","id":"PMC_38177673","title":"Reduction of DHHC5-mediated beclin 1 S-palmitoylation underlies autophagy decline in aging.","date":"2024","source":"Nature structural & molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/38177673","citation_count":35,"is_preprint":false},{"pmid":"31547976","id":"PMC_31547976","title":"DHHC5 Mediates β-Adrenergic Signaling in Cardiomyocytes by Targeting Gα Proteins.","date":"2019","source":"Biophysical journal","url":"https://pubmed.ncbi.nlm.nih.gov/31547976","citation_count":35,"is_preprint":false},{"pmid":"38250049","id":"PMC_38250049","title":"Palmitoylation of PKCδ by ZDHHC5 in hypothalamic microglia presents as a therapeutic target for fatty liver disease.","date":"2024","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/38250049","citation_count":32,"is_preprint":false},{"pmid":"32958558","id":"PMC_32958558","title":"The palmitoyl acyltransferases ZDHHC5 and ZDHHC8 are uniquely present in DRG axons and control retrograde signaling via the Gp130/JAK/STAT3 pathway.","date":"2020","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/32958558","citation_count":32,"is_preprint":false},{"pmid":"38233791","id":"PMC_38233791","title":"ZDHHC5-mediated S-palmitoylation of FAK promotes its membrane localization and epithelial-mesenchymal transition in glioma.","date":"2024","source":"Cell communication and signaling : CCS","url":"https://pubmed.ncbi.nlm.nih.gov/38233791","citation_count":30,"is_preprint":false},{"pmid":"33392180","id":"PMC_33392180","title":"Circ-ZDHHC5 Accelerates Esophageal Squamous Cell Carcinoma Progression in vitro via miR-217/ZEB1 Axis.","date":"2020","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/33392180","citation_count":30,"is_preprint":false},{"pmid":"36774350","id":"PMC_36774350","title":"Repositioning Lomitapide to block ZDHHC5-dependant palmitoylation on SSTR5 leads to anti-proliferation effect in preclinical pancreatic cancer models.","date":"2023","source":"Cell death discovery","url":"https://pubmed.ncbi.nlm.nih.gov/36774350","citation_count":27,"is_preprint":false},{"pmid":"21820437","id":"PMC_21820437","title":"Somatostatin receptor 5 is palmitoylated by the interacting ZDHHC5 palmitoyltransferase.","date":"2011","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/21820437","citation_count":27,"is_preprint":false},{"pmid":"33415776","id":"PMC_33415776","title":"Regulation and function of the palmitoyl-acyltransferase ZDHHC5.","date":"2021","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/33415776","citation_count":26,"is_preprint":false},{"pmid":"30610172","id":"PMC_30610172","title":"Anthrax toxin requires ZDHHC5-mediated palmitoylation of its surface-processing host enzymes.","date":"2019","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/30610172","citation_count":26,"is_preprint":false},{"pmid":"34961524","id":"PMC_34961524","title":"The interactions of ZDHHC5/GOLGA7 with SARS-CoV-2 spike (S) protein and their effects on S protein's subcellular localization, palmitoylation and pseudovirus entry.","date":"2021","source":"Virology journal","url":"https://pubmed.ncbi.nlm.nih.gov/34961524","citation_count":25,"is_preprint":false},{"pmid":"34724258","id":"PMC_34724258","title":"DHHC5 facilitates oligodendrocyte development by palmitoylating and activating STAT3.","date":"2021","source":"Glia","url":"https://pubmed.ncbi.nlm.nih.gov/34724258","citation_count":23,"is_preprint":false},{"pmid":"37163424","id":"PMC_37163424","title":"C-Phycocyanin Ameliorates the Senescence of Mesenchymal Stem Cells through ZDHHC5-Mediated Autophagy via PI3K/AKT/mTOR Pathway.","date":"2023","source":"Aging and disease","url":"https://pubmed.ncbi.nlm.nih.gov/37163424","citation_count":22,"is_preprint":false},{"pmid":"36277202","id":"PMC_36277202","title":"Dynamic but discordant alterations in zDHHC5 expression and palmitoylation of its substrates in cardiac pathologies.","date":"2022","source":"Frontiers in physiology","url":"https://pubmed.ncbi.nlm.nih.gov/36277202","citation_count":21,"is_preprint":false},{"pmid":"35927718","id":"PMC_35927718","title":"Propofol enhances stem-like properties of glioma via GABAAR-dependent Src modulation of ZDHHC5-EZH2 palmitoylation mechanism.","date":"2022","source":"Stem cell research & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/35927718","citation_count":20,"is_preprint":false},{"pmid":"29185452","id":"PMC_29185452","title":"DHHC5-mediated palmitoylation of S1P receptor subtype 1 determines G-protein coupling.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/29185452","citation_count":19,"is_preprint":false},{"pmid":"28739689","id":"PMC_28739689","title":"LXR Activation Down-regulates Lipid Raft Markers FLOT2 and DHHC5 in MCF-7 Breast Cancer Cells.","date":"2017","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/28739689","citation_count":15,"is_preprint":false},{"pmid":"38266744","id":"PMC_38266744","title":"SMPDL3B is palmitoylated and stabilized by ZDHHC5, and its silencing aggravates diabetic retinopathy of db/db mice: Activation of NLRP3/NF-κB pathway.","date":"2024","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/38266744","citation_count":12,"is_preprint":false},{"pmid":"39929269","id":"PMC_39929269","title":"Neonatal sevoflurane exposures inhibits DHHC5-mediated palmitoylation of TfR1 in oligodendrocytes, leading to hypomyelination and neurological impairments.","date":"2025","source":"Journal of advanced research","url":"https://pubmed.ncbi.nlm.nih.gov/39929269","citation_count":11,"is_preprint":false},{"pmid":"35597282","id":"PMC_35597282","title":"Palmitoylation of the small GTPase Cdc42 by DHHC5 modulates spine formation and gene transcription.","date":"2022","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/35597282","citation_count":11,"is_preprint":false},{"pmid":"32202303","id":"PMC_32202303","title":"miR-96-5p enhances cell proliferation and invasion via targeted regulation of ZDHHC5 in gastric cancer.","date":"2020","source":"Bioscience reports","url":"https://pubmed.ncbi.nlm.nih.gov/32202303","citation_count":9,"is_preprint":false},{"pmid":"40109131","id":"PMC_40109131","title":"Protocatechuic Acid Reduces Liver Fatty Acid Uptake in HFD-Fed Mice Associated With the Inhibition of DHHC5-Mediated CD36 Palmitoylation.","date":"2025","source":"Molecular nutrition & food research","url":"https://pubmed.ncbi.nlm.nih.gov/40109131","citation_count":8,"is_preprint":false},{"pmid":"36762613","id":"PMC_36762613","title":"Cell cycle-dependent palmitoylation of protocadherin 7 by ZDHHC5 promotes successful cytokinesis.","date":"2023","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/36762613","citation_count":7,"is_preprint":false},{"pmid":"37557176","id":"PMC_37557176","title":"S-acylation of the Wnt receptor Frizzled-5 by zDHHC5 controls its cellular localization and synaptogenic activity in the rodent hippocampus.","date":"2023","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/37557176","citation_count":6,"is_preprint":false},{"pmid":"41182177","id":"PMC_41182177","title":"ZDHHC5 deficiency impairs spermatogenesis and causes male infertility in mice.","date":"2025","source":"Reproduction (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/41182177","citation_count":1,"is_preprint":false},{"pmid":"42134490","id":"PMC_42134490","title":"ZDHHC5-mediated BRAF palmitoylation activates the MAPK pathway and drives cholangiocarcinoma progression.","date":"2026","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/42134490","citation_count":1,"is_preprint":false},{"pmid":"40589731","id":"PMC_40589731","title":"DHHC5 regulates lacteal function and intestinal lipid absorption by maintaining VEGFR2 localization in lipid rafts.","date":"2025","source":"Life metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/40589731","citation_count":1,"is_preprint":false},{"pmid":"41086885","id":"PMC_41086885","title":"The emerging role of palmitoyl acyltransferase zDHHC5 in health and disease: A review.","date":"2025","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/41086885","citation_count":1,"is_preprint":false},{"pmid":"40898659","id":"PMC_40898659","title":"ZDHHC5 Mediates Immune Dysregulation Driving Generalized Anxiety Disorder Risk.","date":"2025","source":"Brain and behavior","url":"https://pubmed.ncbi.nlm.nih.gov/40898659","citation_count":1,"is_preprint":false},{"pmid":"41179831","id":"PMC_41179831","title":"Amelioration of intervertebral disc degeneration using engineered extracellular vesicle-delivered ZDHHC5 via inhibiting PANoptosis.","date":"2025","source":"Journal of tissue engineering","url":"https://pubmed.ncbi.nlm.nih.gov/41179831","citation_count":0,"is_preprint":false},{"pmid":"42071838","id":"PMC_42071838","title":"DNA methylation-regulated ZDHHC5 and PPT1 in the pathogenesis of osteoporosis.","date":"2026","source":"Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/42071838","citation_count":0,"is_preprint":false},{"pmid":"42098785","id":"PMC_42098785","title":"ZDHHC5: a pivotal palmitoyltransferase orchestrating signaling networks - unraveling mechanisms and therapeutic horizons.","date":"2026","source":"Biomarker research","url":"https://pubmed.ncbi.nlm.nih.gov/42098785","citation_count":0,"is_preprint":false},{"pmid":"41388152","id":"PMC_41388152","title":"ZDHHC5 and ZDHHC14 promote depression via the mediation of double-negative T cells.","date":"2025","source":"Mammalian genome : official journal of the International Mammalian Genome Society","url":"https://pubmed.ncbi.nlm.nih.gov/41388152","citation_count":0,"is_preprint":false},{"pmid":"42260863","id":"PMC_42260863","title":"miR-130a-3p regulates the occurrence and progression of lung squamous cell carcinoma by activating ZDHHC5 and immune cell-related pathways.","date":"2026","source":"Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/42260863","citation_count":0,"is_preprint":false},{"pmid":"41890956","id":"PMC_41890956","title":"Metabolic orchestration of NOD1 signaling by AMPK-mediated phosphorylation of ZDHHC5.","date":"2026","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/41890956","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.07.24.666230","title":"Palmitoylation-dependent activation of NADK promotes NADP  <sup>+</sup>  synthesis and tumorigenesis","date":"2025-07-27","source":"bioRxiv","url":"https://doi.org/10.1101/2025.07.24.666230","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.04.28.651057","title":"Functional Dissection of the Zdhhc5-GOLGA7 Protein Palmitoylation Complex","date":"2025-04-29","source":"bioRxiv","url":"https://doi.org/10.1101/2025.04.28.651057","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.05.26.656092","title":"Syk activation during FcγR-mediated phagocytosis involves both Syk palmitoylation and desulfenylation","date":"2025-05-27","source":"bioRxiv","url":"https://doi.org/10.1101/2025.05.26.656092","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":26749,"output_tokens":9908,"usd":0.114433,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":19968,"output_tokens":4862,"usd":0.110695,"stage2_stop_reason":"end_turn"},"total_usd":0.225128,"stage1_batch_id":"msgbatch_01UT1AahFPCAyn2uVweS7A9z","stage2_batch_id":"msgbatch_01HuKWtKNT4Pqj1bc8eEP5RV","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2010,\n      \"finding\": \"DHHC5 co-immunoprecipitates with PSD-95 in neurons; this interaction is mediated through binding of the C-terminus of DHHC5 to the PDZ3 domain of PSD-95. DHHC5 is highly enriched in post-synaptic density preparations and localizes to CA3 and dentate gyrus of the hippocampus.\",\n      \"method\": \"Co-immunoprecipitation, subcellular fractionation (PSD preparation), immunohistochemistry\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP with domain mapping, single lab, multiple methods\",\n      \"pmids\": [\"20178993\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"DHHC5 palmitoylates flotillin-2; palmitoylation and oligomerization of flotillin-2 are abolished in DHHC5-deficient neuronal stem cells, and overexpression of DHHC5 in COS-1 cells markedly stimulates flotillin-2 palmitoylation, establishing a direct enzyme-substrate relationship.\",\n      \"method\": \"Bioorthogonal palmitate analog (17-octadecynoic acid) metabolic labeling, quantitative proteomics, overexpression in COS-1 cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (metabolic labeling + overexpression rescue), single lab\",\n      \"pmids\": [\"22081607\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"DHHC5 down-regulation is triggered within minutes of growth factor withdrawal from neural stem cells (a signal for neuronal differentiation), and this degradation is partially prevented by inhibitors of ubiquitin-mediated proteolysis, indicating DHHC5 protein levels are regulated by ubiquitin-dependent degradation in response to differentiation signals.\",\n      \"method\": \"Growth factor withdrawal experiments, proteasome inhibitor treatment, time-course protein level analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct pharmacological intervention with defined readout, single lab\",\n      \"pmids\": [\"22081607\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"ZDHHC5 palmitoylates somatostatin receptor 5 (SSTR5); ZDHHC5 interacts with the membrane-proximal helix 8 of SSTR5 and co-localizes with it at the plasma membrane. Overexpression of ZDHHC5 increases SSTR5 palmitoylation and knockdown decreases it, identifying ZDHHC5 as the first palmitoyltransferase for a GPCR.\",\n      \"method\": \"Ras recruitment interaction screen, co-immunoprecipitation, siRNA knockdown, palmitoylation assay\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus gain- and loss-of-function palmitoylation assays, single lab\",\n      \"pmids\": [\"21820437\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"DHHC5 (together with DHHC8) palmitoylates GRIP1b via a PDZ ligand unique to DHHC5/8. Palmitoylated GRIP1b is targeted to trafficking endosomes, where it links endosomes to kinesin motors and accelerates AMPA receptor recycling. GRIP1b palmitoylation turnover rate is among the highest reported.\",\n      \"method\": \"Binding assays, palmitoylation assays, live imaging of dendritic endosomes, AMPA-R recycling assays\",\n      \"journal\": \"Neuron\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (binding, palmitoylation, trafficking, functional receptor assay), replicated with domain mutants\",\n      \"pmids\": [\"22325201\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"DHHC5 palmitoylates phospholemman (PLM) at cysteines C40 and C42 (C40 being the principal site) in cardiac ventricular myocytes. DHHC5 localizes to caveolin-enriched cell surface microdomains and co-immunoprecipitates with PLM. Substrate recruitment requires a ~120 amino acid region of the DHHC5 C-tail immediately after the fourth transmembrane domain, not its PDZ-binding motif. PLM palmitoylation at C40, but not C42, is required for PLM-mediated inhibition of the Na+/K+ ATPase.\",\n      \"method\": \"Co-immunoprecipitation, overexpression and siRNA silencing, site-directed mutagenesis (C40A, C42A), Na pump activity assay, subcellular fractionation\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — mutagenesis of both enzyme (C-tail deletion) and substrate (C40A/C42A) combined with functional Na pump assay, multiple orthogonal methods\",\n      \"pmids\": [\"25422474\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"DHHC5 transiently palmitoylates δ-catenin after enhanced synaptic activity, promoting δ-catenin-cadherin interactions at synapses. This palmitoylation is required for activity-induced stabilization of N-cadherin at synapses, enlargement of postsynaptic spines, insertion of GluA1/GluA2 into the synaptic membrane, and increased mEPSC amplitude. Context-dependent fear conditioning in mice increased δ-catenin palmitoylation and δ-catenin-cadherin associations at hippocampal synapses.\",\n      \"method\": \"Palmitoylation assay, co-immunoprecipitation, spine morphology imaging, electrophysiology (mEPSC), in vivo fear conditioning\",\n      \"journal\": \"Nature neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (biochemistry, imaging, electrophysiology, in vivo), replicated across cell and animal contexts\",\n      \"pmids\": [\"24562000\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Under basal conditions, DHHC5 is bound to PSD-95 and Fyn kinase at the synaptic membrane; Fyn-mediated phosphorylation of a tyrosine residue within the endocytic motif of DHHC5 stabilizes it at the membrane. Neuronal activity disrupts the DHHC5/PSD-95/Fyn complex, causing DHHC5 endocytosis and translocation to dendritic shafts where it encounters and palmitoylates its substrate δ-catenin, after which both return to spines.\",\n      \"method\": \"Live-cell imaging, co-immunoprecipitation, pharmacological inhibition of Fyn, endocytosis assays, palmitoylation assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (live imaging, Co-IP, pharmacological perturbation, palmitoylation), mechanistic model confirmed by functional readouts\",\n      \"pmids\": [\"26334723\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Catalytic activity of DHHC5 is required for NSCLC cell proliferation, colony formation, invasion, and tumor xenograft growth; rescue with wild-type but not catalytically inactive DHHC5 confirms the dependence on enzymatic activity.\",\n      \"method\": \"siRNA/shRNA knockdown, lentiviral re-expression of WT vs. catalytic mutant DHHC5, in vitro proliferation/invasion assays, xenograft mouse models\",\n      \"journal\": \"Molecular cancer research : MCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — catalytic mutant rescue experiment in both cell culture and in vivo, single lab\",\n      \"pmids\": [\"25573953\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"ZDHHC5 palmitoylates EZH2 in glioma cells; mutant p53 transcriptionally upregulates ZDHHC5 (together with NF-Y), which alters EZH2 palmitoylation and phosphorylation status, promoting glioma stem-like cell self-renewal and tumorigenicity.\",\n      \"method\": \"Co-immunoprecipitation, palmitoylation assay, ChIP/reporter assay for transcriptional regulation, glioma stem cell self-renewal assays\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple methods (palmitoylation assay, transcriptional mechanism, functional stem cell assay), single lab\",\n      \"pmids\": [\"28775165\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"DHHC5 palmitoylates the S1P receptor subtype 1 (S1P1R) at the plasma membrane, which is prerequisite for Gi protein coupling. Upon receptor stimulation and internalization, S1P1R is depalmitoylated. FTY720-P-induced endocytosed S1P1R vesicles subsequently associate with DHHC5, enabling persistent intracellular Gi signaling.\",\n      \"method\": \"Palmitoylation assay, co-immunoprecipitation, subcellular localization imaging, cAMP/Gi signaling assays, receptor internalization assays\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — palmitoylation assay linked to functional G-protein coupling readout, multiple methods, single lab\",\n      \"pmids\": [\"29185452\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"DHHC4 and DHHC5 palmitoylate CD36 at different subcellular localizations to control its plasma membrane localization and fatty acid uptake activity. Depletion of DHHC5 disrupts CD36-dependent fatty acid uptake in cells, and adipose-specific Dhhc5 knockout mice show decreased fatty acid uptake in adipose tissue and develop hypothermia upon cold exposure.\",\n      \"method\": \"Palmitoylation assay, siRNA/shRNA knockdown, fatty acid uptake assay, conditional knockout mouse model, cold exposure phenotype\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — cell-based and in vivo knockout with multiple mechanistic readouts (palmitoylation, localization, function), replicated with two enzymes\",\n      \"pmids\": [\"30605677\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"ZDHHC5 forms a catalytically active protein S-acyltransferase complex with the accessory subunit GOLGA7. The ZDHHC5-GOLGA7 complex is mutually stabilizing and localizes to the plasma membrane. This complex is required for CIL56-induced nonapoptotic cell death and is involved in retrograde protein trafficking from the plasma membrane.\",\n      \"method\": \"CRISPR genetic screen, co-immunoprecipitation, subcellular localization imaging, cell death assays with catalytic mutants\",\n      \"journal\": \"Cell chemical biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR screen plus Co-IP and catalytic mutant validation, single lab\",\n      \"pmids\": [\"31631010\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"DHHC5 palmitoylates the proprotein convertases Furin and PC7. ZDHHC5-mediated palmitoylation promotes Furin/PC7 association with plasma membrane microdomains, which is required for cleavage of anthrax toxin protective antigen (but does not inhibit protease activity per se). ZDHHC5 also affects homeostasis and trafficking of Furin/PC7 through the endocytic/recycling pathway.\",\n      \"method\": \"Palmitoylation assay, subcellular fractionation (lipid raft), siRNA knockdown, anthrax toxin processing assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — palmitoylation assay plus functional toxin processing readout, single lab, multiple methods\",\n      \"pmids\": [\"30610172\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"DHHC5 binds to and palmitoylates Golga7b. Palmitoylation of Golga7b prevents clathrin-mediated endocytosis of DHHC5 and stabilizes it at the plasma membrane. Desmoglein-2 and plakophilin-3 are substrates of DHHC5, and DHHC5/Golga7b are required for desmoglein-2 plasma membrane localization, desmosomal patterning, and cell adhesion.\",\n      \"method\": \"Co-immunoprecipitation, palmitoylation assay, siRNA knockdown, fluorescence imaging, proteomic analysis of complexes, cell adhesion assays\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (Co-IP, palmitoylation assay, proteomics, imaging, functional adhesion assay), multiple substrates identified\",\n      \"pmids\": [\"31402609\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"DHHC5 mediates β-adrenergic stimulation-dependent palmitoylation of Gαs and Gαi in cardiomyocytes. An in vitro palmitoylation assay with purified components established Gαs and Gαi as direct DHHC5 substrates. The C-terminal tail of DHHC5 itself is palmitoylated in response to stimulation, which is important for DHHC5 dynamic localization and function at the plasma membrane.\",\n      \"method\": \"In vitro palmitoylation assay with purified components, siRNA knockdown, cAMP measurement, contractility assay, subcellular localization imaging\",\n      \"journal\": \"Biophysical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with purified components plus functional cellular readouts, single lab\",\n      \"pmids\": [\"31547976\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Among all mammalian PATs, only ZDHHC5 and ZDHHC8 are enriched in DRG axons. Both are required for Gp130/JAK/STAT3 (but not DLK/JNK) axon-to-soma retrograde signaling after injury. ZDHHC5 and ZDHHC8 palmitoylate Gp130 in co-transfected cells; shRNA knockdown reduces Gp130 palmitoylation and surface expression in DRG neurons.\",\n      \"method\": \"Immunofluorescence localization screen of all PATs in DRG neurons, shRNA knockdown, palmitoylation assay, STAT3/JNK signaling assays, co-transfection palmitoylation assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — comprehensive PAT localization screen plus palmitoylation and signaling assays, single lab\",\n      \"pmids\": [\"32958558\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DHHC5 palmitoylates STAT3 in oligodendrocytes; STAT3 is the primary DHHC5 substrate in these cells. DHHC5 ablation reduces STAT3 palmitoylation and suppresses STAT3 phosphorylation/activation, inhibiting transcription of myelin-related and anti-apoptosis genes and suppressing myelination and remyelination.\",\n      \"method\": \"Conditional Zdhhc5 knockout, palmitoylation assay, STAT3 phosphorylation western blot, myelination histology, gene expression analysis\",\n      \"journal\": \"Glia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional KO with mechanistic substrate identification and functional myelination readout, single lab\",\n      \"pmids\": [\"34724258\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ZDHHC5 and GOLGA7 interact with the SARS-CoV-2 spike protein and are required for efficient SARS-CoV-2 pseudovirus entry into A549 and HeLa cells. ZDHHC5/GOLGA7 knockout had no significant effect on S protein subcellular localization or palmitoylation, but significantly decreased pseudovirus entry.\",\n      \"method\": \"CRISPR-Cas9 knockout, co-immunoprecipitation, ABE palmitoylation assay, pseudovirus entry luciferase assay, fluorescence microscopy\",\n      \"journal\": \"Virology journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR KO plus multiple assays, single lab; mechanism of entry effect not fully resolved\",\n      \"pmids\": [\"34961524\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"DHHC5 palmitoylates the brain-specific isoform Cdc42-palm; DHHC5 physically interacts with Cdc42-palm. A dominant-negative DHHC5 mutant or DHHC5 knockdown decreases Cdc42-palm palmitoylation and plasma membrane localization, leading to reduced Cdc42-mediated gene transcription and spine formation in hippocampal neurons.\",\n      \"method\": \"Co-immunoprecipitation, palmitoylation assay, dominant-negative and shRNA approaches, fluorescence imaging, spine morphology analysis, transcription reporter assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus loss-of-function with multiple cellular readouts, single lab\",\n      \"pmids\": [\"35597282\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ZDHHC5 palmitoylates NLRP3 at the LRR domain. This palmitoylation promotes NLRP3 oligomerization, NLRP3-NEK7 interaction, and ASC aggregate formation, leading to caspase-1 activation, IL-1β/18 release, and GSDMD cleavage. ABHD17A acts as the depalmitoylase for NLRP3. Zdhhc5-/- mice show defective NLRP3 inflammasome activation in vivo.\",\n      \"method\": \"Palmitoylation assay, siRNA/KO, co-immunoprecipitation (NLRP3-NEK7), ASC speck imaging, caspase-1 activation assay, IL-1β/18 ELISA, GSDMD cleavage, Zdhhc5 KO mouse model\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (palmitoylation assay, protein-protein interaction, inflammasome assembly, in vivo KO), replicated in human cells and mice\",\n      \"pmids\": [\"38092000\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ZDHHC5 palmitoylates FAK at C456, promoting FAK membrane localization and phosphorylation (activation). ZDHHC5 knockdown abolishes FAK palmitoylation and membrane distribution, reducing cell proliferation, invasion, and EMT in glioblastoma cells and tumor growth in intracranial xenografts.\",\n      \"method\": \"Acyl-PEG exchange assay, metabolic incorporation assay, subcellular fractionation/imaging, siRNA knockdown, 2-BP inhibition, CCK8/invasion/colony assays, intracranial xenograft\",\n      \"journal\": \"Cell communication and signaling : CCS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — palmitoylation site identified by metabolic assay plus functional in vitro and in vivo readouts, single lab\",\n      \"pmids\": [\"38233791\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ZDHHC5 palmitoylates protocadherin 7 (PCDH7) in a cell cycle-dependent manner. PCDH7 and ZDHHC5 colocalize at the mitotic cell surface and translocate together to the cleavage furrow. PCDH7 localization at the cell surface depends on ZDHHC5 palmitoylation activity. Loss of PCDH7 increases multinucleated cells, prolongs mitosis, and reduces active RhoA and phospho-myosin at the cleavage furrow.\",\n      \"method\": \"Co-immunoprecipitation, palmitoylation assay, live-cell imaging, siRNA knockdown, RhoA activity assay, phospho-myosin immunofluorescence\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus palmitoylation-dependent localization and functional cytokinesis readouts, single lab\",\n      \"pmids\": [\"36762613\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ZDHHC5 palmitoylates SSTR5 and lomitapide, an FDA-approved drug, acts as an inhibitor of ZDHHC5, blocking palmitoylation of SSTR5 and reducing pancreatic cancer cell growth in vitro and in vivo.\",\n      \"method\": \"Palmitoylation assay, drug treatment (lomitapide), cell proliferation assays, in vivo xenograft\",\n      \"journal\": \"Cell death discovery\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — pharmacological inhibitor study with functional readout; mechanism of ZDHHC5 inhibition by lomitapide not fully characterized, single lab\",\n      \"pmids\": [\"36774350\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Frizzled-5 (Fz5) is S-acylated on three C-terminal cysteines by zDHHC5. S-acylation is essential for Fz5 localization to the cell surface, axons, and presynaptic sites; S-acylation-deficient Fz5 is internalized faster and fails to activate canonical and divergent canonical Wnt pathways. S-acylation levels of Fz5 are regulated by neuronal activity, and S-acylation-deficient Fz5 fails to induce presynaptic assembly in vivo.\",\n      \"method\": \"Palmitoylation assay, mutagenesis of acylation sites, live imaging, internalization assay, Wnt pathway reporter assay, in vivo presynaptic assembly assay\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — mutagenesis of acylation sites combined with functional pathway and in vivo synaptogenesis assays, multiple orthogonal methods\",\n      \"pmids\": [\"37557176\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"DHHC5 mediates palmitoylation of beclin 1, which promotes formation of the ATG14L-containing class III PI3K complex I (PI3KC3-C1) and its lipid kinase activity, by enhancing hydrophobic interactions between beclin 1 and adapter proteins ATG14L and VPS15. DHHC5 deficiency in neurons exaggerates neurodegeneration in Alzheimer's disease mouse models in an autophagy-dependent manner.\",\n      \"method\": \"Palmitoylation assay, co-immunoprecipitation, PI3K lipid kinase activity assay, Zdhhc5 KO neuronal models, autophagy flux analysis, AD mouse models\",\n      \"journal\": \"Nature structural & molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — enzymatic activity assay (PI3K) linked directly to palmitoylation, multiple orthogonal methods, disease-relevant in vivo models\",\n      \"pmids\": [\"38177673\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZDHHC5 palmitoylates PKCδ in hypothalamic microglia; an antimalarial drug (artemether) blocks ZDHHC5-PKCδ binding to inhibit PKCδ palmitoylation, suppressing downstream neuroinflammation signaling and improving hepatic lipid metabolism via TRH/thyroid hormone neuroendocrine axis.\",\n      \"method\": \"Biotin-labeled chemical probe pulldown, co-immunoprecipitation, palmitoylation assay, siRNA knockdown, neuroendocrine assays, high-fat diet mouse model\",\n      \"journal\": \"Theranostics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — chemical probe target ID plus Co-IP, palmitoylation assay, and in vivo KD, single lab\",\n      \"pmids\": [\"38250049\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZDHHC5 palmitoylates SMPDL3B, and high glucose exposure increases ZDHHC5-SMPDL3B interaction. Palmitoylation by ZDHHC5 stabilizes SMPDL3B protein; inhibiting palmitoylation accelerates SMPDL3B degradation, while inhibiting depalmitoylation decreases its turnover rate in retinal endothelial cells.\",\n      \"method\": \"Co-immunoprecipitation, palmitoylation inhibitor (2-BP) and depalmitoylation inhibitor (palmostatin B) treatment, protein stability assay\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — co-IP and pharmacological inhibitors without direct palmitoylation site identification or reconstitution, single lab\",\n      \"pmids\": [\"38266744\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZDHHC5 palmitoylates BRAF at Cys194/195 in cholangiocarcinoma cells, promoting BRAF membrane localization and protein stabilization. This activates ERK/MAPK signaling. Knockdown of ZDHHC5 or mutation of BRAF palmitoylation sites inhibits ERK signaling and cholangiocarcinoma growth.\",\n      \"method\": \"Palmitoylation assay, site-directed mutagenesis (C194/195A), ERK western blot, siRNA knockdown, cell proliferation assay\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — palmitoylation site mutagenesis plus functional signaling and growth assays, single lab\",\n      \"pmids\": [\"42134490\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"DHHC5 palmitoylates transferrin receptor 1 (TfR1) at C98 in oligodendrocytes, which is required for regulation of TfR1 endocytosis. In neonatal sevoflurane-exposed mice, DHHC5 is specifically downregulated in oligodendrocytes, reducing TfR1 palmitoylation, enhancing TfR1 endocytosis, causing iron accumulation and ferroptosis, leading to hypomyelination.\",\n      \"method\": \"Acyl-resin assisted capture assay, co-immunoprecipitation, specific oligodendrocyte DHHC5 overexpression (Pdgfrα-CreERT), single-cell RNA sequencing, TfR1 endocytosis assay\",\n      \"journal\": \"Journal of advanced research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — palmitoylation site identified plus conditional rescue in specific cell type in vivo, single lab\",\n      \"pmids\": [\"39929269\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ZDHHC5 mediates IFT81 palmitoylation in round spermatids; ZDHHC5 predominantly localizes to the Golgi apparatus of round spermatids (steps 2-8). Zdhhc5 KO male mice are completely infertile with defective spermatogenesis, reduced sperm motility, and sperm tail malformations linked to defective IFT81 regulation.\",\n      \"method\": \"CRISPR/Cas9 knockout, immunofluorescence localization, palmitoylation assay, sperm morphology/motility analysis, IVF, ICSI rescue\",\n      \"journal\": \"Reproduction (Cambridge, England)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO with specific cellular localization and substrate identification plus functional rescue experiment (ICSI), single lab\",\n      \"pmids\": [\"41182177\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"DHHC5 palmitoylates CRYBG1 (an actin-binding protein) in lymphatic endothelial cells; CRYBG1 palmitoylation is required for VEGFR2 lipid raft localization, which promotes VEGFR2 signaling and intestinal lymphatic integrity. Dhhc5 intestinal/LEC-specific knockout mice show impaired intestinal lipid absorption and resistance to diet-induced obesity.\",\n      \"method\": \"Conditional Dhhc5 knockout, palmitoylation assay, lipid raft fractionation, VEGFR2 signaling assay, lipid absorption assay\",\n      \"journal\": \"Life metabolism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional KO with mechanistic substrate identification and functional lipid absorption readout, single lab\",\n      \"pmids\": [\"40589731\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"AMPK phosphorylates ZDHHC5 at Ser296 and Ser380, which weakens ZDHHC5 association with GOLGA7B and promotes ZDHHC5 displacement from the plasma membrane, thereby reducing NOD1 palmitoylation, NOD1 membrane localization, and downstream NOD1 innate immune signaling. Conversely, NOD1 agonist C12-iE-DAP suppresses AMPK activity to stabilize ZDHHC5 at the membrane.\",\n      \"method\": \"Biochemical phosphorylation assay, site-directed mutagenesis (Ser296/380), co-immunoprecipitation, palmitoylation assay, NOD1 membrane fractionation, NF-κB signaling assay\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phosphorylation site mutagenesis plus functional NOD1 signaling readout, multiple methods, single lab\",\n      \"pmids\": [\"41890956\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ZDHHC5 palmitoylates NADK at Cys22, Cys23, and Cys26 within its amino-terminal domain, stimulating NADK kinase activity by relieving an autoinhibitory function of the amino terminus and promoting NADP+ synthesis. Zdhhc5-/- mice show defective NADK palmitoylation and NADP+ production.\",\n      \"method\": \"In vitro palmitoylation assay, site-directed mutagenesis, NADK kinase activity assay, Zdhhc5 KO mouse model, metabolomics (NADP+ measurement)\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — in vitro assay with mutagenesis and in vivo KO, but preprint (not peer-reviewed), single lab\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"The ZDHHC5-GOLGA7 complex structure was determined by cryo-EM. Key conserved residues in both ZDHHC5 and GOLGA7 required for complex formation were identified by mutagenesis; disruption of these residues abolished CIL56-induced nonapoptotic cell death.\",\n      \"method\": \"Cryo-EM structure determination, biochemical purification, homology modeling, mutagenesis, cell death functional assays\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — cryo-EM structure with mutagenesis validation, but preprint (not peer-reviewed), single lab\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"DHHC5 palmitoylates Syk on a single cysteine residue during FcγR-mediated phagocytosis in macrophages. Syk palmitoylation is required for Syk localization to the phagocytic cup, Syk phosphorylation/activation, Cdc42 recruitment, F-actin polymerization, and phagocytosis.\",\n      \"method\": \"S-acylation assay, site-directed mutagenesis, fluorescence imaging of phagocytic cup, Syk phosphorylation western blot, phagocytosis assay\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — multiple mechanistic readouts with mutagenesis, but preprint (not peer-reviewed), single lab\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ZDHHC5 palmitoylates YBX1 (via PANoptosis-related mechanism in intervertebral disc cells), and ZDHHC5-enriched extracellular vesicles modulate ZBP1 transcription through competitive inhibition of YBX1 phosphorylation via palmitoylation.\",\n      \"method\": \"Transcriptomics, scRNA-seq, co-immunoprecipitation, palmitoylation assay, extracellular vesicle delivery, transcriptional reporter assay\",\n      \"journal\": \"Journal of tissue engineering\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — mechanism proposed from EV-delivery experiments; palmitoylation of YBX1 by ZDHHC5 not directly confirmed by reconstitution, single lab\",\n      \"pmids\": [\"41179831\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ZDHHC5 is a plasma membrane-localized DHHC-family palmitoyl acyltransferase that S-palmitoylates a broad range of substrates—including CD36, GRIP1b, δ-catenin, PLM, flotillin-2, NLRP3, Gαs/Gαi, beclin 1, EZH2, STAT3, FAK, BRAF, S1P1R, Fz5, SSTR5, NOD1, Cdc42, Syk, PCDH7, Furin/PC7, Gp130, desmoglein-2, IFT81, CRYBG1, TfR1, and NADK—thereby controlling their membrane localization, stability, activity, and interactions; ZDHHC5 activity is itself regulated by Fyn-mediated tyrosine phosphorylation of its endocytic motif (controlling activity-dependent neuronal trafficking), AMPK-mediated phosphorylation at Ser296/380 (coupling metabolic state to innate immune signaling), palmitoylation of its own C-terminal tail, and complex formation with accessory proteins GOLGA7/Golga7b that stabilize it at the plasma membrane; through these mechanisms ZDHHC5 governs diverse processes including synaptic plasticity, AMPA receptor recycling, cardiac sodium pump regulation, β-adrenergic signaling, NLRP3 inflammasome assembly, autophagy, myelination, fatty acid uptake, cytokinesis, axonal retrograde signaling, and cell adhesion.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ZDHHC5 is a plasma-membrane DHHC-family protein S-acyltransferase that palmitoylates a broad set of substrates to control their membrane localization, stability, and activity, thereby governing synaptic plasticity, innate immunity, metabolism, and cell adhesion [#4, #5, #20]. At neuronal synapses it is held at the membrane through a complex with PSD-95 and Fyn, in which Fyn-mediated tyrosine phosphorylation of the ZDHHC5 endocytic motif stabilizes it; neuronal activity disrupts this complex and drives ZDHHC5 endocytosis to dendritic shafts where it transiently palmitoylates \\u03b4-catenin to stabilize cadherin and remodel spines [#6, #7], and it likewise palmitoylates GRIP1b to accelerate AMPA-receptor recycling [#4] and Cdc42-palm to support spine formation [#19]. Its enzymatic activity is required for substrate effects, demonstrated by catalytically inactive rescue failures across multiple systems [#5, #8]. ZDHHC5 is stabilized at the plasma membrane by the accessory subunit GOLGA7, with which it forms a mutually stabilizing, catalytically active complex, and by palmitoylation of GOLGA7b that blocks its clathrin-mediated endocytosis [#12, #14]; its own C-terminal tail is also palmitoylated in a stimulus-dependent manner to tune membrane dynamics [#15]. Through palmitoylation of NLRP3 it promotes inflammasome oligomerization, NLRP3-NEK7 interaction, and caspase-1 activation, with ABHD17A serving as the opposing depalmitoylase [#20], and AMPK phosphorylation at Ser296/Ser380 weakens its GOLGA7B association to dampen NOD1 signaling, coupling metabolic state to innate immunity [#32]. ZDHHC5 also palmitoylates beclin 1 to assemble the ATG14L-containing PI3KC3-C1 autophagy complex [#25], CD36 to drive fatty acid uptake [#11], desmoglein-2 to pattern desmosomes and mediate adhesion [#14], and signaling effectors including BRAF, FAK, STAT3, and Gp130 that link it to MAPK/ERK, STAT3, and retrograde axonal signaling and to tumor and myelination phenotypes [#16, #17, #21, #28].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"Established where and with what ZDHHC5 associates in the brain, anchoring it physically to the postsynaptic density rather than treating it as a diffuse membrane enzyme.\",\n      \"evidence\": \"Reciprocal Co-IP with domain mapping, PSD fractionation, and immunohistochemistry in neurons\",\n      \"pmids\": [\"20178993\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not identify catalytic substrates\", \"Functional consequence of the PSD-95 interaction unresolved at this stage\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Identified the first concrete substrates (flotillin-2, SSTR5) and showed ZDHHC5 protein levels are themselves controlled by ubiquitin-dependent degradation, establishing it as a regulated enzyme with defined targets.\",\n      \"evidence\": \"Metabolic palmitate labeling, overexpression rescue, Ras-recruitment interaction screen, and proteasome inhibitor treatment in neural stem cells and COS-1 cells\",\n      \"pmids\": [\"22081607\", \"21820437\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Palmitoylation site residues not mapped\", \"Mechanism of differentiation-triggered degradation not defined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Connected ZDHHC5 catalysis to a defined neuronal trafficking output by showing GRIP1b palmitoylation routes endosomes to kinesin motors and accelerates AMPA-receptor recycling.\",\n      \"evidence\": \"Binding and palmitoylation assays, live imaging of dendritic endosomes, AMPA-R recycling assays\",\n      \"pmids\": [\"22325201\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Redundancy with DHHC8 not fully separated\", \"Endogenous regulation of this turnover not addressed\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Resolved how ZDHHC5 selects substrates and linked its activity to physiology, mapping a substrate-recruitment region in the C-tail and showing site-specific PLM palmitoylation controls Na+/K+ ATPase regulation, plus activity-dependent \\u03b4-catenin palmitoylation driving synaptic plasticity.\",\n      \"evidence\": \"Enzyme C-tail deletion and substrate C40A/C42A mutagenesis with Na pump assay in cardiomyocytes; palmitoylation, spine imaging, mEPSC, and in vivo fear conditioning\",\n      \"pmids\": [\"25422474\", \"24562000\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of C-tail substrate recruitment unknown\", \"How activity is transduced to palmitoylation not yet mechanistic\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Defined the trafficking control logic of ZDHHC5 itself, showing a Fyn/PSD-95-phosphorylation switch governs its membrane retention versus endocytic relocation to reach substrates, and demonstrated strict dependence on catalytic activity for oncogenic phenotypes.\",\n      \"evidence\": \"Live imaging, Co-IP, Fyn pharmacological inhibition, and endocytosis assays; catalytic-mutant rescue in NSCLC cells and xenografts\",\n      \"pmids\": [\"26334723\", \"25573953\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the cancer-relevant substrate(s) not defined in the NSCLC study\", \"Generality of the Fyn switch to non-neuronal cells unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Extended ZDHHC5 into GPCR signaling persistence and tumor biology, showing S1P1R palmitoylation enables Gi coupling and that mutant p53 transcriptionally upregulates ZDHHC5 to alter EZH2 in glioma stem cells.\",\n      \"evidence\": \"Palmitoylation assays with Gi signaling and internalization readouts; ChIP/reporter assays and stem-cell self-renewal assays\",\n      \"pmids\": [\"29185452\", \"28775165\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"EZH2 palmitoylation sites not mapped\", \"Direct enzyme-substrate proof for EZH2 limited to Co-IP/palmitoylation\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Established the GOLGA7/Golga7b accessory module that stabilizes ZDHHC5 at the membrane and broadly expanded its substrate range to metabolism (CD36), adhesion (desmoglein-2), secretion (Furin/PC7), and G-protein/self-palmitoylation, including in vivo knockout phenotypes.\",\n      \"evidence\": \"CRISPR screen, Co-IP, conditional Dhhc5 KO mice with cold-exposure and fatty-acid-uptake phenotypes, in vitro palmitoylation with purified G\\u03b1 components, adhesion and toxin-processing assays\",\n      \"pmids\": [\"31631010\", \"30605677\", \"31402609\", \"30610172\", \"31547976\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the ZDHHC5-GOLGA7 complex not yet resolved at this stage\", \"Rules governing which substrate is engaged in which compartment unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrated cell-type-specific dominant substrates, with STAT3 as the principal oligodendrocyte target controlling myelination through palmitoylation-dependent STAT3 activation.\",\n      \"evidence\": \"Conditional Zdhhc5 knockout, palmitoylation assay, STAT3 phosphorylation, and myelination histology\",\n      \"pmids\": [\"34724258\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"STAT3 palmitoylation site not specified\", \"How palmitoylation couples to STAT3 phosphorylation mechanistically unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Positioned ZDHHC5 as a central node in inflammasome assembly and cytokinesis and as a druggable cancer target, showing LRR-domain NLRP3 palmitoylation (opposed by ABHD17A) drives caspase-1 activation, FAK and BRAF palmitoylation drive tumor signaling, and PCDH7 palmitoylation supports cleavage-furrow function.\",\n      \"evidence\": \"Palmitoylation and site-directed mutagenesis, NLRP3-NEK7 Co-IP, ASC speck and caspase-1 assays, Zdhhc5 KO mice, RhoA/phospho-myosin and xenograft assays, lomitapide inhibitor study\",\n      \"pmids\": [\"38092000\", \"38233791\", \"36762613\", \"36774350\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Lomitapide's direct mode of ZDHHC5 inhibition not characterized (Low-confidence)\", \"Whether a single ZDHHC5 conformation handles such diverse substrates is unresolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Linked ZDHHC5 to autophagy machinery assembly and additional metabolic/neuroinflammatory axes, showing beclin 1 palmitoylation builds the ATG14L PI3KC3-C1 complex with disease relevance in Alzheimer's models.\",\n      \"evidence\": \"Palmitoylation assay, PI3K lipid-kinase activity assay, Zdhhc5 KO neuronal and AD mouse models; PKC\\u03b4 and SMPDL3B substrate studies\",\n      \"pmids\": [\"38177673\", \"38250049\", \"38266744\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"SMPDL3B palmitoylation site and direct catalysis not established (Low-confidence)\", \"Beclin 1 palmitoylation site within the corpus not detailed\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Provided the first ZDHHC5-GOLGA7 cryo-EM structure and a metabolic regulatory loop (AMPK phosphorylation at Ser296/380 weakening GOLGA7B binding to dampen NOD1 signaling), plus new substrates spanning iron handling, lymphatic lipid uptake, NADK activation, spermatogenesis, and phagocytosis.\",\n      \"evidence\": \"Cryo-EM and mutagenesis (preprint), AMPK phosphosite mutagenesis with NOD1 signaling, conditional KO and KO mouse models, in vitro NADK kinase assay (preprint), Syk phagocytosis assay (preprint)\",\n      \"pmids\": [\"41890956\", \"39929269\", \"40589731\", \"41182177\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Key structural and NADK/Syk findings remain in preprint form\", \"A unifying model for compartment- and stimulus-specific substrate selection is still absent\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unknown how a single ZDHHC5 enzyme achieves selectivity across its very large substrate set and how its localization, GOLGA7/GOLGA7b association, phosphorylation, and self-palmitoylation are integrated to direct it to the right substrate in the right compartment.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No comprehensive substrate-recognition code defined\", \"Limited structural insight into catalysis-substrate engagement\", \"Cross-talk between the multiple regulatory inputs not reconstituted\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [4, 5, 11, 14, 20, 25]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [1, 5, 15, 24]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [20, 25, 33]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [3, 5, 12, 14, 15]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [30]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [10, 12]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [20, 32]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [25]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [10, 16, 28]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [11, 31]},\n      {\"term_id\": \"R-HSA-112316\", \"supporting_discovery_ids\": [4, 6, 7]}\n    ],\n    \"complexes\": [\n      \"ZDHHC5-GOLGA7 S-acyltransferase complex\",\n      \"DHHC5/PSD-95/Fyn synaptic complex\"\n    ],\n    \"partners\": [\n      \"GOLGA7\",\n      \"GOLGA7B\",\n      \"PSD-95\",\n      \"FYN\",\n      \"NLRP3\",\n      \"CD36\",\n      \"NEK7\",\n      \"BECN1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}