{"gene":"ATP2B3","run_date":"2026-06-09T22:02:44","timeline":{"discoveries":[{"year":2013,"finding":"Somatic hotspot mutations in ATP2B3 (encoding plasma membrane Ca2+-ATPase 3) were identified in aldosterone-producing adenomas (APAs). Electrophysiological ex vivo studies on primary adrenal adenoma cells provided evidence for inappropriate depolarization of cells with ATP2B3 alterations, consistent with loss of normal Ca2+ pump function driving autonomous aldosterone secretion.","method":"Exome sequencing of APAs, electrophysiological ex vivo studies on primary adrenal adenoma cells","journal":"Nature genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ex vivo electrophysiology plus exome sequencing, single study, functional data primarily shown for ATP1A1 mutants with electrophysiology extended to ATP2B3-mutant cells","pmids":["23416519"],"is_preprint":false},{"year":2013,"finding":"Somatic ATP2B3 mutations in APAs lead to upregulation of CYP11B2 (aldosterone synthase) gene expression. Overexpression of ATP2B3 mutants in HAC15 adrenal cells resulted in increased CYP11B2 and its transcriptional regulator NR4A2 expression.","method":"Sanger sequencing, overexpression in HAC15 adrenal cells, gene expression analysis (CYP11B2, NR4A2)","journal":"Hypertension (Dallas, Tex. : 1979)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — cell-based overexpression with defined molecular readout (CYP11B2, NR4A2 upregulation), single lab","pmids":["24082052"],"is_preprint":false},{"year":2016,"finding":"The APA-associated ATP2B3 Leu425_Val426del mutation causes: (1) reduced Ca2+ export due to loss of physiological pump function; (2) elevated basal intracellular Ca2+ and increased Ca2+ influx; (3) a Na+-dependent inward current that strongly depolarizes the plasma membrane. These effects collectively increase aldosterone synthase (CYP11B2) mRNA expression and enhance aldosterone production.","method":"Ca2+ measurements (live-cell imaging), electrophysiology (patch-clamp), mRNA expression analysis, aldosterone assay in NCI-H295R and HEK-293 cells overexpressing mutant ATP2B3","journal":"Endocrinology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (Ca2+ imaging, patch-clamp, aldosterone quantification, mRNA), functional mechanism clearly delineated in two cell lines","pmids":["27035656"],"is_preprint":false},{"year":2007,"finding":"The 14-3-3epsilon protein interacts with PMCA3 (ATP2B3) in a phosphorylation-independent manner, and this interaction inhibits PMCA3 pump activity. Co-expression of 14-3-3epsilon with PMCA3 in CHO cells substantially decreased the ability of cells to restore basal Ca2+ concentration following an InsP3-induced Ca2+ transient. The 14-3-3zeta isoform also interacted with PMCA3.","method":"Two-hybrid assay, co-immunoprecipitation, GST pull-down, aequorin-based Ca2+ measurements in CHO cells","journal":"Cell calcium","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal binding confirmed by multiple methods (two-hybrid, co-IP, pull-down) plus functional readout (Ca2+ homeostasis assay), multiple PMCA isoforms tested as controls","pmids":["18029012"],"is_preprint":false},{"year":1995,"finding":"PMCA3 protein is expressed in rat brain neurons and is localized primarily in cell processes (granule cell processes in cerebellum, dendritic fields in hippocampus CA1), with the translated protein having an observed molecular mass of ~135 kDa as predicted from molecular cloning.","method":"In situ hybridization, immunoblot analysis with anti-peptide antibodies, immunohistochemistry, co-localization with GAP-43 marker","journal":"Brain research. Molecular brain research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct protein detection by immunoblot and immunohistochemistry with subcellular localization in brain, single lab","pmids":["7770003"],"is_preprint":false},{"year":1994,"finding":"ATP2B3 (PMCA isoform 3) was mapped to human chromosome Xq28 by fluorescence in situ hybridization (FISH), somatic cell hybrid analysis, and genetic linkage analysis.","method":"FISH, somatic cell hybrid analysis, genetic linkage analysis","journal":"Cytogenetics and cell genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — chromosomal localization established by three independent genomic methods in a single study","pmids":["8187550"],"is_preprint":false},{"year":2017,"finding":"A novel PMCA3 G733R substitution in the catalytic P-domain impairs the pump's ability to control cellular Ca2+ handling under both basal and stimulated conditions. Homology modeling and molecular dynamics revealed that the mutated residue maintains the correct 3D configuration of the local pump structure.","method":"Biochemical Ca2+ handling assays (live-cell), homology modeling, molecular dynamics simulation","journal":"Biochimica et biophysica acta. Molecular basis of disease","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — functional Ca2+ assay in cells plus computational structural analysis, single lab, no in vitro reconstitution","pmids":["28807751"],"is_preprint":false},{"year":2022,"finding":"The ataxia-linked PMCA3 E1081Q mutation (immediately upstream of the calmodulin-binding domain) shows a splicing variant-dependent effect: in the full-length b variant it abolishes the pump's capacity to reduce sub-plasma membrane [Ca2+], while in the truncated a variant it selectively increases Ca2+ extrusion activity in sub-plasma membrane microdomains.","method":"Live-cell Ca2+ microdomain measurements, expression of variant-specific PMCA3 constructs, biochemical/molecular characterization","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional Ca2+ measurement in sub-PM microdomains with two splice variants tested, single lab","pmids":["36207321"],"is_preprint":false},{"year":2012,"finding":"Downregulation of PMCA3 in PC12 cells accelerates dibutyryl-cAMP-induced neuronal differentiation and results in longer neurites, altered expression of voltage-dependent Ca2+ channels (VDCCs), and compensatory upregulation of PMCA1, PMCA4, and SERCA, indicating that PMCA3 participates in the control of neuronal Ca2+ handling machinery and differentiation.","method":"Antisense-mediated stable knockdown in PC12 cells, morphological analysis, Ca2+ influx assays, immunoblot for Ca2+ handling proteins","journal":"Cell calcium","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — stable KD with defined cellular and molecular phenotype, single lab, multiple readouts","pmids":["22921123"],"is_preprint":false},{"year":2023,"finding":"ATP2B3 knockdown in HT-22 cells alleviates erastin-induced ferroptosis by acting through the P62-KEAP1-NRF2-HO-1 pathway: knockdown reversed upregulation of P62, NRF2, HO-1, NQO1 and downregulation of KEAP1 induced by erastin, and reduced ROS production. HO-1 overexpression reversed the protective effect of ATP2B3 inhibition, confirming pathway placement.","method":"siRNA knockdown, TMT-based proteomics, cell viability assay, ROS measurement, epistasis by NRF2 KD/P62 inhibition/KEAP1 OE, HO-1 overexpression rescue","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with multiple pathway components tested, single lab, neuronal cell line","pmids":["37298147"],"is_preprint":false},{"year":2021,"finding":"A novel ATP2B3 K416_F418delinsN somatic mutation causes a functional defect: HAC15 adrenal cells transfected with this mutant showed increased CYP11B2 expression and increased aldosterone production.","method":"Sanger sequencing of APA tissue, transfection of mutant gene into HAC15 cells, CYP11B2 expression analysis, aldosterone production assay","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — cell-based functional assay with single mutant, single lab","pmids":["34572956"],"is_preprint":false},{"year":2011,"finding":"Suppression of PMCA3 in GH3 cells increased GAD65 expression, indicating that PMCA3 activity modulates GABA synthesis machinery in pituitary cells.","method":"Antisense-mediated knockdown of PMCA3 in GH3 cells, enzyme activity assays (GAD, GABA-T, SSADH), immunoblot","journal":"Biochemical and biophysical research communications","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single KD approach with limited mechanistic depth","pmids":["21798237"],"is_preprint":false}],"current_model":"ATP2B3 (PMCA3) is a plasma membrane Ca2+-ATPase that actively exports Ca2+ from cells to maintain cytosolic Ca2+ homeostasis, particularly in neurons and adrenal cells; loss-of-function somatic mutations in its transmembrane domain cause reduced Ca2+ extrusion, membrane depolarization, elevated intracellular Ca2+, and aberrant activation of aldosterone synthase (CYP11B2) in adrenal adenomas, while in neurons its activity is inhibited by 14-3-3 proteins (epsilon and zeta isoforms binding phosphorylation-independently) and germline mutations in its P-domain or calmodulin-binding region impair sub-plasma membrane Ca2+ microdomain regulation and are linked to cerebellar ataxia; additionally, ATP2B3 participates in ferroptosis resistance via the P62-KEAP1-NRF2-HO-1 pathway."},"narrative":{"mechanistic_narrative":"ATP2B3 (PMCA3) is a plasma membrane Ca2+-ATPase that actively extrudes cytosolic Ca2+ to maintain Ca2+ homeostasis, with prominent roles in neuronal and adrenal cells [PMID:27035656, PMID:7770003]. In neurons it is expressed primarily in cell processes, where it shapes Ca2+ handling: its loss accelerates differentiation and triggers compensatory upregulation of other Ca2+ transporters (PMCA1, PMCA4, SERCA), and it regulates Ca2+ specifically in sub-plasma membrane microdomains [PMID:7770003, PMID:22921123, PMID:36207321]. Pump activity is negatively regulated by 14-3-3epsilon (and 14-3-3zeta) through phosphorylation-independent binding that blunts the cell's ability to restore basal Ca2+ after a transient [PMID:18029012]. Loss-of-function somatic mutations clustered in the transmembrane domain — including in-frame deletions/indels such as Leu425_Val426del and K416_F418delinsN — impair Ca2+ export, elevate basal intracellular Ca2+, and produce a Na+-dependent depolarizing current; this drives upregulation of aldosterone synthase (CYP11B2) and its regulator NR4A2, accounting for autonomous aldosterone secretion in aldosterone-producing adenomas [PMID:23416519, PMID:24082052, PMID:27035656, PMID:34572956]. Germline mutations in the catalytic P-domain (G733R) and adjacent to the calmodulin-binding region (E1081Q) impair Ca2+ handling, with the E1081Q effect dependent on the b splice variant, linking PMCA3 dysfunction to cerebellar ataxia [PMID:28807751, PMID:36207321]. Knockdown of ATP2B3 also confers resistance to erastin-induced ferroptosis through the P62-KEAP1-NRF2-HO-1 axis [PMID:37298147].","teleology":[{"year":1994,"claim":"Establishing the genomic locus of ATP2B3 placed the gene on the X chromosome, a prerequisite for later linkage of germline mutations to X-linked phenotypes.","evidence":"FISH, somatic cell hybrid analysis, and genetic linkage mapping to Xq28","pmids":["8187550"],"confidence":"Medium","gaps":["Does not address protein function or tissue expression","No mechanistic data"]},{"year":1995,"claim":"Determining where PMCA3 protein resides answered whether it is positioned to control local neuronal Ca2+, showing enrichment in neuronal cell processes.","evidence":"In situ hybridization, immunoblot, and immunohistochemistry with co-localization to neuronal processes in rat brain","pmids":["7770003"],"confidence":"Medium","gaps":["Subcellular microdomain resolution not addressed","Functional consequence of process localization not tested"]},{"year":2007,"claim":"Identifying a regulator of PMCA3 activity revealed that 14-3-3epsilon/zeta bind the pump phosphorylation-independently and inhibit Ca2+ clearance, defining a post-translational brake on the pump.","evidence":"Two-hybrid, co-IP, GST pull-down, and aequorin Ca2+ measurements in CHO cells","pmids":["18029012"],"confidence":"High","gaps":["Binding site on PMCA3 not mapped","Physiological context of inhibition in neurons not tested"]},{"year":2011,"claim":"Linking PMCA3 to neurotransmitter synthesis machinery indicated its Ca2+ handling influences GABA synthesis enzymes in pituitary cells.","evidence":"Antisense knockdown in GH3 cells with GAD/GABA-T/SSADH enzyme assays and immunoblot","pmids":["21798237"],"confidence":"Low","gaps":["Single lab, single knockdown approach with limited mechanistic depth","Direct link between Ca2+ extrusion and GAD65 not established","Not confirmed in vivo"]},{"year":2012,"claim":"Knockdown phenotyping addressed PMCA3's role in neuronal Ca2+ machinery and differentiation, showing it shapes neurite outgrowth and is buffered by compensatory transporters.","evidence":"Antisense stable knockdown in PC12 cells with morphology, Ca2+ influx assays, and immunoblot for Ca2+ handling proteins","pmids":["22921123"],"confidence":"Medium","gaps":["Compensation by PMCA1/4/SERCA complicates interpretation of direct role","Differentiation mechanism downstream of Ca2+ not resolved"]},{"year":2013,"claim":"Discovery of recurrent somatic ATP2B3 mutations in aldosterone-producing adenomas, with depolarization in mutant cells, established a loss-of-pump mechanism driving autonomous aldosterone secretion.","evidence":"Exome/Sanger sequencing of APAs, ex vivo electrophysiology, and overexpression in HAC15 cells with CYP11B2/NR4A2 readout","pmids":["23416519","24082052"],"confidence":"Medium","gaps":["Detailed biophysical mechanism of depolarization not resolved in 2013","Causality of Ca2+ changes for CYP11B2 induction inferred"]},{"year":2016,"claim":"Mechanistic dissection of the Leu425_Val426del mutant resolved how transmembrane-domain loss-of-function causes disease: reduced Ca2+ export, elevated basal Ca2+, increased influx, and a Na+-dependent depolarizing current driving CYP11B2/aldosterone.","evidence":"Ca2+ imaging, patch-clamp, mRNA analysis, and aldosterone assay in NCI-H295R and HEK-293 cells","pmids":["27035656"],"confidence":"High","gaps":["Molecular basis of the aberrant Na+-dependent current not structurally defined","In vivo validation absent"]},{"year":2017,"claim":"Characterizing a P-domain mutation (G733R) extended PMCA3 dysfunction to the catalytic core, showing impaired Ca2+ handling without gross structural perturbation.","evidence":"Live-cell Ca2+ assays plus homology modeling and molecular dynamics","pmids":["28807751"],"confidence":"Medium","gaps":["No in vitro reconstitution of catalytic defect","Genotype–phenotype link to ataxia inferred from cell data"]},{"year":2021,"claim":"A new APA mutation (K416_F418delinsN) confirmed the recurrent functional logic of ATP2B3 loss-of-function in driving CYP11B2 and aldosterone.","evidence":"Sanger sequencing of APA tissue and transfection into HAC15 cells with CYP11B2 and aldosterone readouts","pmids":["34572956"],"confidence":"Medium","gaps":["Single mutant, no electrophysiology","Ca2+ measurements not reported for this variant"]},{"year":2022,"claim":"Analysis of the ataxia-linked E1081Q mutation revealed a splice-variant-dependent effect on sub-plasma membrane Ca2+ microdomains, refining how calmodulin-region mutations alter local Ca2+ control.","evidence":"Live-cell sub-PM Ca2+ microdomain measurements with variant-specific PMCA3 constructs","pmids":["36207321"],"confidence":"Medium","gaps":["Mechanism by which splice variant determines opposite effects unresolved","In vivo neuronal consequence not tested"]},{"year":2023,"claim":"Placing ATP2B3 in a ferroptosis pathway showed its knockdown protects neuronal cells via P62-KEAP1-NRF2-HO-1 signaling, broadening its role beyond Ca2+ pumping.","evidence":"siRNA knockdown, TMT proteomics, viability and ROS assays, and epistasis/rescue with NRF2/P62/KEAP1/HO-1 in HT-22 cells","pmids":["37298147"],"confidence":"Medium","gaps":["Direct molecular connection between Ca2+ extrusion and NRF2 pathway not established","Single cell line"]},{"year":null,"claim":"How PMCA3 Ca2+ extrusion is mechanistically coupled to its downstream regulatory outputs — CYP11B2 induction in adrenal cells, ataxia in neurons, and the ferroptosis pathway — remains unresolved at the molecular level.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No high-resolution structure of human PMCA3 or its mutants","14-3-3 binding site and integration with mutation effects unmapped","Mechanism linking Ca2+ handling to NRF2/ferroptosis signaling unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[2]},{"term_id":"GO:0005215","term_label":"transporter activity","supporting_discovery_ids":[2,7,8]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[2,6]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[2,4,7]}],"pathway":[],"complexes":[],"partners":["YWHAE","YWHAZ"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q16720","full_name":"Plasma membrane calcium-transporting ATPase 3","aliases":["Plasma membrane calcium ATPase isoform 3","Plasma membrane calcium pump isoform 3"],"length_aa":1220,"mass_kda":134.2,"function":"ATP-driven Ca(2+) ion pump involved in the maintenance of basal intracellular Ca(2+) levels at the presynaptic terminals (PubMed:18029012, PubMed:22912398, PubMed:25953895, PubMed:27035656). Uses ATP as an energy source to transport cytosolic Ca(2+) ions across the plasma membrane to the extracellular compartment (PubMed:25953895, PubMed:27035656). May counter-transport protons, but the mechanism and the stoichiometry of this Ca(2+)/H(+) exchange remains to be established (By similarity)","subcellular_location":"Cell membrane; Presynaptic cell membrane","url":"https://www.uniprot.org/uniprotkb/Q16720/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ATP2B3","classification":"Not Classified","n_dependent_lines":5,"n_total_lines":1208,"dependency_fraction":0.0041390728476821195},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"ATP2B1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/ATP2B3","total_profiled":1310},"omim":[{"mim_id":"302500","title":"SPINOCEREBELLAR ATAXIA, X-LINKED 1; SCAX1","url":"https://www.omim.org/entry/302500"},{"mim_id":"300014","title":"ATPase, Ca(2+)-TRANSPORTING, PLASMA MEMBRANE, 3; ATP2B3","url":"https://www.omim.org/entry/300014"},{"mim_id":"182310","title":"ATPase, Na+/K+ TRANSPORTING, ALPHA-1 POLYPEPTIDE; ATP1A1","url":"https://www.omim.org/entry/182310"},{"mim_id":"108733","title":"ATPase, Ca(2+)-TRANSPORTING, PLASMA MEMBRANE, 2; ATP2B2","url":"https://www.omim.org/entry/108733"},{"mim_id":"108732","title":"ATPase, Ca(2+)-TRANSPORTING, PLASMA MEMBRANE, 4; ATP2B4","url":"https://www.omim.org/entry/108732"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Plasma membrane","reliability":"Approved"}],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"choroid plexus","ntpm":106.2}],"url":"https://www.proteinatlas.org/search/ATP2B3"},"hgnc":{"alias_symbol":["PMCA3","CFAP39"],"prev_symbol":["SCAX1","CLA2"]},"alphafold":{"accession":"Q16720","domains":[{"cath_id":"2.70.150.10","chopping":"30-88_211-289","consensus_level":"medium","plddt":85.6199,"start":30,"end":289},{"cath_id":"1.20.1110.10","chopping":"358-444_840-1066","consensus_level":"high","plddt":84.5335,"start":358,"end":1066},{"cath_id":"3.40.50.1000","chopping":"453-478_684-830","consensus_level":"high","plddt":86.8229,"start":453,"end":830},{"cath_id":"3.40.1110.10","chopping":"483-679","consensus_level":"high","plddt":86.569,"start":483,"end":679}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q16720","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q16720-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q16720-F1-predicted_aligned_error_v6.png","plddt_mean":74.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ATP2B3","jax_strain_url":"https://www.jax.org/strain/search?query=ATP2B3"},"sequence":{"accession":"Q16720","fasta_url":"https://rest.uniprot.org/uniprotkb/Q16720.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q16720/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q16720"}},"corpus_meta":[{"pmid":"23416519","id":"PMC_23416519","title":"Somatic mutations in ATP1A1 and ATP2B3 lead to aldosterone-producing adenomas and secondary hypertension.","date":"2013","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23416519","citation_count":448,"is_preprint":false},{"pmid":"24082052","id":"PMC_24082052","title":"Somatic ATP1A1, ATP2B3, and KCNJ5 mutations in aldosterone-producing adenomas.","date":"2013","source":"Hypertension (Dallas, Tex. : 1979)","url":"https://pubmed.ncbi.nlm.nih.gov/24082052","citation_count":135,"is_preprint":false},{"pmid":"17071747","id":"PMC_17071747","title":"CLA-1 and its splicing variant CLA-2 mediate bacterial adhesion and cytosolic bacterial invasion in mammalian cells.","date":"2006","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/17071747","citation_count":57,"is_preprint":false},{"pmid":"27035656","id":"PMC_27035656","title":"Cellular Pathophysiology of an Adrenal Adenoma-Associated Mutant of the Plasma Membrane Ca(2+)-ATPase ATP2B3.","date":"2016","source":"Endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/27035656","citation_count":52,"is_preprint":false},{"pmid":"7770003","id":"PMC_7770003","title":"Localization of the plasma membrane Ca(2+)-ATPase isoform PMCA3 in rat cerebellum, choroid plexus and hippocampus.","date":"1995","source":"Brain research. Molecular brain research","url":"https://pubmed.ncbi.nlm.nih.gov/7770003","citation_count":45,"is_preprint":false},{"pmid":"8187550","id":"PMC_8187550","title":"Localization of two genes encoding plasma membrane Ca2+ ATPases isoforms 2 (ATP2B2) and 3 (ATP2B3) to human chromosomes 3p26-->p25 and Xq28, respectively.","date":"1994","source":"Cytogenetics and cell genetics","url":"https://pubmed.ncbi.nlm.nih.gov/8187550","citation_count":32,"is_preprint":false},{"pmid":"18029012","id":"PMC_18029012","title":"Inhibitory interaction of the 14-3-3 proteins with ubiquitous (PMCA1) and tissue-specific (PMCA3) isoforms of the plasma membrane Ca2+ pump.","date":"2007","source":"Cell calcium","url":"https://pubmed.ncbi.nlm.nih.gov/18029012","citation_count":26,"is_preprint":false},{"pmid":"22921123","id":"PMC_22921123","title":"Downregulation of PMCA2 or PMCA3 reorganizes Ca(2+) handling systems in differentiating PC12 cells.","date":"2012","source":"Cell calcium","url":"https://pubmed.ncbi.nlm.nih.gov/22921123","citation_count":25,"is_preprint":false},{"pmid":"15277554","id":"PMC_15277554","title":"Characterization and expression of plasma membrane Ca2+ ATPase (PMCA3) in the crayfish Procambarus clarkii antennal gland during molting.","date":"2004","source":"The Journal of experimental biology","url":"https://pubmed.ncbi.nlm.nih.gov/15277554","citation_count":24,"is_preprint":false},{"pmid":"23773993","id":"PMC_23773993","title":"Exome sequencing identification of a GJB1 missense mutation in a kindred with X-linked spinocerebellar ataxia (SCA-X1).","date":"2013","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23773993","citation_count":18,"is_preprint":false},{"pmid":"2584925","id":"PMC_2584925","title":"Analysis of a novel VHS107 haplotype in CLA-2 and WSA mice. Evidence for gene conversion among IgVH genes in outbred populations.","date":"1989","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/2584925","citation_count":14,"is_preprint":false},{"pmid":"28807751","id":"PMC_28807751","title":"A novel PMCA3 mutation in an ataxic patient with hypomorphic phosphomannomutase 2 (PMM2) heterozygote mutations: Biochemical characterization of the pump defect.","date":"2017","source":"Biochimica et biophysica acta. Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/28807751","citation_count":12,"is_preprint":false},{"pmid":"37298147","id":"PMC_37298147","title":"ATP2B3 Inhibition Alleviates Erastin-Induced Ferroptosis in HT-22 Cells through the P62-KEAP1-NRF2-HO-1 Pathway.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/37298147","citation_count":6,"is_preprint":false},{"pmid":"23007728","id":"PMC_23007728","title":"Ectopic expression of a maize calreticulin mitigates calcium deficiency-like disorders in sCAX1-expressing tobacco and tomato.","date":"2012","source":"Plant molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/23007728","citation_count":5,"is_preprint":false},{"pmid":"36207321","id":"PMC_36207321","title":"The ataxia-linked E1081Q mutation affects the sub-plasma membrane Ca2+-microdomains by tuning PMCA3 activity.","date":"2022","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/36207321","citation_count":5,"is_preprint":false},{"pmid":"26481629","id":"PMC_26481629","title":"A Novel Somatic Deletion Mutation of ATP2B3 in Aldosterone-Producing Adenoma.","date":"2015","source":"Endocrine pathology","url":"https://pubmed.ncbi.nlm.nih.gov/26481629","citation_count":5,"is_preprint":false},{"pmid":"21798237","id":"PMC_21798237","title":"GABA-shunt enzymes activity in GH3 cells with reduced level of PMCA2 or PMCA3 isoform.","date":"2011","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/21798237","citation_count":4,"is_preprint":false},{"pmid":"34572956","id":"PMC_34572956","title":"Characteristics of a Novel ATP2B3 K416_F418delinsN Mutation in a Classical Aldosterone-Producing Adenoma.","date":"2021","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/34572956","citation_count":1,"is_preprint":false},{"pmid":"36081280","id":"PMC_36081280","title":"Expression, purification and structure determination of the chlorinase ClA2.","date":"2022","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/36081280","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11970,"output_tokens":3004,"usd":0.040485,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10478,"output_tokens":3630,"usd":0.07157,"stage2_stop_reason":"end_turn"},"total_usd":0.112055,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2013,\n      \"finding\": \"Somatic hotspot mutations in ATP2B3 (encoding plasma membrane Ca2+-ATPase 3) were identified in aldosterone-producing adenomas (APAs). Electrophysiological ex vivo studies on primary adrenal adenoma cells provided evidence for inappropriate depolarization of cells with ATP2B3 alterations, consistent with loss of normal Ca2+ pump function driving autonomous aldosterone secretion.\",\n      \"method\": \"Exome sequencing of APAs, electrophysiological ex vivo studies on primary adrenal adenoma cells\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ex vivo electrophysiology plus exome sequencing, single study, functional data primarily shown for ATP1A1 mutants with electrophysiology extended to ATP2B3-mutant cells\",\n      \"pmids\": [\"23416519\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Somatic ATP2B3 mutations in APAs lead to upregulation of CYP11B2 (aldosterone synthase) gene expression. Overexpression of ATP2B3 mutants in HAC15 adrenal cells resulted in increased CYP11B2 and its transcriptional regulator NR4A2 expression.\",\n      \"method\": \"Sanger sequencing, overexpression in HAC15 adrenal cells, gene expression analysis (CYP11B2, NR4A2)\",\n      \"journal\": \"Hypertension (Dallas, Tex. : 1979)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — cell-based overexpression with defined molecular readout (CYP11B2, NR4A2 upregulation), single lab\",\n      \"pmids\": [\"24082052\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The APA-associated ATP2B3 Leu425_Val426del mutation causes: (1) reduced Ca2+ export due to loss of physiological pump function; (2) elevated basal intracellular Ca2+ and increased Ca2+ influx; (3) a Na+-dependent inward current that strongly depolarizes the plasma membrane. These effects collectively increase aldosterone synthase (CYP11B2) mRNA expression and enhance aldosterone production.\",\n      \"method\": \"Ca2+ measurements (live-cell imaging), electrophysiology (patch-clamp), mRNA expression analysis, aldosterone assay in NCI-H295R and HEK-293 cells overexpressing mutant ATP2B3\",\n      \"journal\": \"Endocrinology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (Ca2+ imaging, patch-clamp, aldosterone quantification, mRNA), functional mechanism clearly delineated in two cell lines\",\n      \"pmids\": [\"27035656\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"The 14-3-3epsilon protein interacts with PMCA3 (ATP2B3) in a phosphorylation-independent manner, and this interaction inhibits PMCA3 pump activity. Co-expression of 14-3-3epsilon with PMCA3 in CHO cells substantially decreased the ability of cells to restore basal Ca2+ concentration following an InsP3-induced Ca2+ transient. The 14-3-3zeta isoform also interacted with PMCA3.\",\n      \"method\": \"Two-hybrid assay, co-immunoprecipitation, GST pull-down, aequorin-based Ca2+ measurements in CHO cells\",\n      \"journal\": \"Cell calcium\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal binding confirmed by multiple methods (two-hybrid, co-IP, pull-down) plus functional readout (Ca2+ homeostasis assay), multiple PMCA isoforms tested as controls\",\n      \"pmids\": [\"18029012\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"PMCA3 protein is expressed in rat brain neurons and is localized primarily in cell processes (granule cell processes in cerebellum, dendritic fields in hippocampus CA1), with the translated protein having an observed molecular mass of ~135 kDa as predicted from molecular cloning.\",\n      \"method\": \"In situ hybridization, immunoblot analysis with anti-peptide antibodies, immunohistochemistry, co-localization with GAP-43 marker\",\n      \"journal\": \"Brain research. Molecular brain research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct protein detection by immunoblot and immunohistochemistry with subcellular localization in brain, single lab\",\n      \"pmids\": [\"7770003\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1994,\n      \"finding\": \"ATP2B3 (PMCA isoform 3) was mapped to human chromosome Xq28 by fluorescence in situ hybridization (FISH), somatic cell hybrid analysis, and genetic linkage analysis.\",\n      \"method\": \"FISH, somatic cell hybrid analysis, genetic linkage analysis\",\n      \"journal\": \"Cytogenetics and cell genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — chromosomal localization established by three independent genomic methods in a single study\",\n      \"pmids\": [\"8187550\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"A novel PMCA3 G733R substitution in the catalytic P-domain impairs the pump's ability to control cellular Ca2+ handling under both basal and stimulated conditions. Homology modeling and molecular dynamics revealed that the mutated residue maintains the correct 3D configuration of the local pump structure.\",\n      \"method\": \"Biochemical Ca2+ handling assays (live-cell), homology modeling, molecular dynamics simulation\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular basis of disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — functional Ca2+ assay in cells plus computational structural analysis, single lab, no in vitro reconstitution\",\n      \"pmids\": [\"28807751\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The ataxia-linked PMCA3 E1081Q mutation (immediately upstream of the calmodulin-binding domain) shows a splicing variant-dependent effect: in the full-length b variant it abolishes the pump's capacity to reduce sub-plasma membrane [Ca2+], while in the truncated a variant it selectively increases Ca2+ extrusion activity in sub-plasma membrane microdomains.\",\n      \"method\": \"Live-cell Ca2+ microdomain measurements, expression of variant-specific PMCA3 constructs, biochemical/molecular characterization\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional Ca2+ measurement in sub-PM microdomains with two splice variants tested, single lab\",\n      \"pmids\": [\"36207321\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Downregulation of PMCA3 in PC12 cells accelerates dibutyryl-cAMP-induced neuronal differentiation and results in longer neurites, altered expression of voltage-dependent Ca2+ channels (VDCCs), and compensatory upregulation of PMCA1, PMCA4, and SERCA, indicating that PMCA3 participates in the control of neuronal Ca2+ handling machinery and differentiation.\",\n      \"method\": \"Antisense-mediated stable knockdown in PC12 cells, morphological analysis, Ca2+ influx assays, immunoblot for Ca2+ handling proteins\",\n      \"journal\": \"Cell calcium\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — stable KD with defined cellular and molecular phenotype, single lab, multiple readouts\",\n      \"pmids\": [\"22921123\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ATP2B3 knockdown in HT-22 cells alleviates erastin-induced ferroptosis by acting through the P62-KEAP1-NRF2-HO-1 pathway: knockdown reversed upregulation of P62, NRF2, HO-1, NQO1 and downregulation of KEAP1 induced by erastin, and reduced ROS production. HO-1 overexpression reversed the protective effect of ATP2B3 inhibition, confirming pathway placement.\",\n      \"method\": \"siRNA knockdown, TMT-based proteomics, cell viability assay, ROS measurement, epistasis by NRF2 KD/P62 inhibition/KEAP1 OE, HO-1 overexpression rescue\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with multiple pathway components tested, single lab, neuronal cell line\",\n      \"pmids\": [\"37298147\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"A novel ATP2B3 K416_F418delinsN somatic mutation causes a functional defect: HAC15 adrenal cells transfected with this mutant showed increased CYP11B2 expression and increased aldosterone production.\",\n      \"method\": \"Sanger sequencing of APA tissue, transfection of mutant gene into HAC15 cells, CYP11B2 expression analysis, aldosterone production assay\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — cell-based functional assay with single mutant, single lab\",\n      \"pmids\": [\"34572956\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Suppression of PMCA3 in GH3 cells increased GAD65 expression, indicating that PMCA3 activity modulates GABA synthesis machinery in pituitary cells.\",\n      \"method\": \"Antisense-mediated knockdown of PMCA3 in GH3 cells, enzyme activity assays (GAD, GABA-T, SSADH), immunoblot\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single KD approach with limited mechanistic depth\",\n      \"pmids\": [\"21798237\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ATP2B3 (PMCA3) is a plasma membrane Ca2+-ATPase that actively exports Ca2+ from cells to maintain cytosolic Ca2+ homeostasis, particularly in neurons and adrenal cells; loss-of-function somatic mutations in its transmembrane domain cause reduced Ca2+ extrusion, membrane depolarization, elevated intracellular Ca2+, and aberrant activation of aldosterone synthase (CYP11B2) in adrenal adenomas, while in neurons its activity is inhibited by 14-3-3 proteins (epsilon and zeta isoforms binding phosphorylation-independently) and germline mutations in its P-domain or calmodulin-binding region impair sub-plasma membrane Ca2+ microdomain regulation and are linked to cerebellar ataxia; additionally, ATP2B3 participates in ferroptosis resistance via the P62-KEAP1-NRF2-HO-1 pathway.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ATP2B3 (PMCA3) is a plasma membrane Ca2+-ATPase that actively extrudes cytosolic Ca2+ to maintain Ca2+ homeostasis, with prominent roles in neuronal and adrenal cells [#2, #4]. In neurons it is expressed primarily in cell processes, where it shapes Ca2+ handling: its loss accelerates differentiation and triggers compensatory upregulation of other Ca2+ transporters (PMCA1, PMCA4, SERCA), and it regulates Ca2+ specifically in sub-plasma membrane microdomains [#4, #8, #7]. Pump activity is negatively regulated by 14-3-3epsilon (and 14-3-3zeta) through phosphorylation-independent binding that blunts the cell's ability to restore basal Ca2+ after a transient [#3]. Loss-of-function somatic mutations clustered in the transmembrane domain — including in-frame deletions/indels such as Leu425_Val426del and K416_F418delinsN — impair Ca2+ export, elevate basal intracellular Ca2+, and produce a Na+-dependent depolarizing current; this drives upregulation of aldosterone synthase (CYP11B2) and its regulator NR4A2, accounting for autonomous aldosterone secretion in aldosterone-producing adenomas [#0, #1, #2, #10]. Germline mutations in the catalytic P-domain (G733R) and adjacent to the calmodulin-binding region (E1081Q) impair Ca2+ handling, with the E1081Q effect dependent on the b splice variant, linking PMCA3 dysfunction to cerebellar ataxia [#6, #7]. Knockdown of ATP2B3 also confers resistance to erastin-induced ferroptosis through the P62-KEAP1-NRF2-HO-1 axis [#9].\",\n  \"teleology\": [\n    {\n      \"year\": 1994,\n      \"claim\": \"Establishing the genomic locus of ATP2B3 placed the gene on the X chromosome, a prerequisite for later linkage of germline mutations to X-linked phenotypes.\",\n      \"evidence\": \"FISH, somatic cell hybrid analysis, and genetic linkage mapping to Xq28\",\n      \"pmids\": [\"8187550\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not address protein function or tissue expression\", \"No mechanistic data\"]\n    },\n    {\n      \"year\": 1995,\n      \"claim\": \"Determining where PMCA3 protein resides answered whether it is positioned to control local neuronal Ca2+, showing enrichment in neuronal cell processes.\",\n      \"evidence\": \"In situ hybridization, immunoblot, and immunohistochemistry with co-localization to neuronal processes in rat brain\",\n      \"pmids\": [\"7770003\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Subcellular microdomain resolution not addressed\", \"Functional consequence of process localization not tested\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Identifying a regulator of PMCA3 activity revealed that 14-3-3epsilon/zeta bind the pump phosphorylation-independently and inhibit Ca2+ clearance, defining a post-translational brake on the pump.\",\n      \"evidence\": \"Two-hybrid, co-IP, GST pull-down, and aequorin Ca2+ measurements in CHO cells\",\n      \"pmids\": [\"18029012\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Binding site on PMCA3 not mapped\", \"Physiological context of inhibition in neurons not tested\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Linking PMCA3 to neurotransmitter synthesis machinery indicated its Ca2+ handling influences GABA synthesis enzymes in pituitary cells.\",\n      \"evidence\": \"Antisense knockdown in GH3 cells with GAD/GABA-T/SSADH enzyme assays and immunoblot\",\n      \"pmids\": [\"21798237\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single lab, single knockdown approach with limited mechanistic depth\", \"Direct link between Ca2+ extrusion and GAD65 not established\", \"Not confirmed in vivo\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Knockdown phenotyping addressed PMCA3's role in neuronal Ca2+ machinery and differentiation, showing it shapes neurite outgrowth and is buffered by compensatory transporters.\",\n      \"evidence\": \"Antisense stable knockdown in PC12 cells with morphology, Ca2+ influx assays, and immunoblot for Ca2+ handling proteins\",\n      \"pmids\": [\"22921123\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Compensation by PMCA1/4/SERCA complicates interpretation of direct role\", \"Differentiation mechanism downstream of Ca2+ not resolved\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Discovery of recurrent somatic ATP2B3 mutations in aldosterone-producing adenomas, with depolarization in mutant cells, established a loss-of-pump mechanism driving autonomous aldosterone secretion.\",\n      \"evidence\": \"Exome/Sanger sequencing of APAs, ex vivo electrophysiology, and overexpression in HAC15 cells with CYP11B2/NR4A2 readout\",\n      \"pmids\": [\"23416519\", \"24082052\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Detailed biophysical mechanism of depolarization not resolved in 2013\", \"Causality of Ca2+ changes for CYP11B2 induction inferred\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Mechanistic dissection of the Leu425_Val426del mutant resolved how transmembrane-domain loss-of-function causes disease: reduced Ca2+ export, elevated basal Ca2+, increased influx, and a Na+-dependent depolarizing current driving CYP11B2/aldosterone.\",\n      \"evidence\": \"Ca2+ imaging, patch-clamp, mRNA analysis, and aldosterone assay in NCI-H295R and HEK-293 cells\",\n      \"pmids\": [\"27035656\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis of the aberrant Na+-dependent current not structurally defined\", \"In vivo validation absent\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Characterizing a P-domain mutation (G733R) extended PMCA3 dysfunction to the catalytic core, showing impaired Ca2+ handling without gross structural perturbation.\",\n      \"evidence\": \"Live-cell Ca2+ assays plus homology modeling and molecular dynamics\",\n      \"pmids\": [\"28807751\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No in vitro reconstitution of catalytic defect\", \"Genotype–phenotype link to ataxia inferred from cell data\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"A new APA mutation (K416_F418delinsN) confirmed the recurrent functional logic of ATP2B3 loss-of-function in driving CYP11B2 and aldosterone.\",\n      \"evidence\": \"Sanger sequencing of APA tissue and transfection into HAC15 cells with CYP11B2 and aldosterone readouts\",\n      \"pmids\": [\"34572956\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single mutant, no electrophysiology\", \"Ca2+ measurements not reported for this variant\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Analysis of the ataxia-linked E1081Q mutation revealed a splice-variant-dependent effect on sub-plasma membrane Ca2+ microdomains, refining how calmodulin-region mutations alter local Ca2+ control.\",\n      \"evidence\": \"Live-cell sub-PM Ca2+ microdomain measurements with variant-specific PMCA3 constructs\",\n      \"pmids\": [\"36207321\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which splice variant determines opposite effects unresolved\", \"In vivo neuronal consequence not tested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Placing ATP2B3 in a ferroptosis pathway showed its knockdown protects neuronal cells via P62-KEAP1-NRF2-HO-1 signaling, broadening its role beyond Ca2+ pumping.\",\n      \"evidence\": \"siRNA knockdown, TMT proteomics, viability and ROS assays, and epistasis/rescue with NRF2/P62/KEAP1/HO-1 in HT-22 cells\",\n      \"pmids\": [\"37298147\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular connection between Ca2+ extrusion and NRF2 pathway not established\", \"Single cell line\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How PMCA3 Ca2+ extrusion is mechanistically coupled to its downstream regulatory outputs — CYP11B2 induction in adrenal cells, ataxia in neurons, and the ferroptosis pathway — remains unresolved at the molecular level.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No high-resolution structure of human PMCA3 or its mutants\", \"14-3-3 binding site and integration with mutation effects unmapped\", \"Mechanism linking Ca2+ handling to NRF2/ferroptosis signaling unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0005215\", \"supporting_discovery_ids\": [2, 7, 8]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [2, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [2, 4, 7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0005509\", \"supporting_discovery_ids\": []}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"YWHAE\",\n      \"YWHAZ\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":6,"faith_pct":83.33333333333333}}