{"gene":"ZDHHC3","run_date":"2026-04-28T23:00:24","timeline":{"discoveries":[{"year":2002,"finding":"GODZ (ZDHHC3) is a Golgi apparatus-specific protein with a DHHC zinc finger domain and four putative transmembrane regions; overexpression in COS7 cells suppressed sorting of glutamate receptor GluRα1 from the Golgi apparatus, implicating ZDHHC3 in membrane protein trafficking.","method":"Overexpression in COS7 cells, subcellular localization by immunofluorescence","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 — single overexpression experiment with defined trafficking phenotype, single lab","pmids":["12163046"],"is_preprint":false},{"year":2004,"finding":"ZDHHC3 (GODZ) was identified as a palmitoyltransferase that palmitoylates the γ2 subunit of GABA(A) receptors via interaction with a conserved 14-amino acid cysteine-rich domain in the large cytoplasmic loop of γ1-3 subunits; GODZ localizes to the Golgi complex in neurons and its coexpression with GABA(A) receptors in heterologous cells results in γ2 palmitoylation in a cytoplasmic loop domain-dependent manner.","method":"SOS-recruitment yeast two-hybrid, coexpression in heterologous cells, metabolic palmitoylation assay, immunofluorescence localization","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (Y2H, palmitoylation assay, domain-dependency mutagenesis), replicated in neurons, highly cited","pmids":["15229235"],"is_preprint":false},{"year":2006,"finding":"ZDHHC3 (GODZ) and its paralog SERZ-β (ZDHHC7) form homomultimers and heteromultimers; GODZ is required for normal synaptic accumulation of GABA(A) receptors, whole-cell and synaptic GABAergic inhibitory function, and GABAergic innervation, but is dispensable for postsynaptic AMPA receptor-mediated glutamatergic transmission; dominant-negative GODZ (C157S) and GODZ-specific RNAi established these requirements.","method":"Coimmunoprecipitation, in vivo cross-linking, dominant-negative expression (GODZ C157S), plasmid-based RNAi, electrophysiology in neurons","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP plus clean KD with defined synaptic phenotype, multiple orthogonal methods, replicated","pmids":["17151279"],"is_preprint":false},{"year":2009,"finding":"ZDHHC3 (GODZ) mediates Ca2+ transport in Xenopus oocytes in a palmitoylation-dependent manner; mutation of the DHHC motif (GODZ-DHHS) or treatment with palmitoylation inhibitor 2-bromopalmitate reduced Ca2+ transport ~80%; a separate V61R mutation abolished Ca2+ transport without affecting palmitoyl acyltransferase activity, indicating dual functions.","method":"Two-electrode voltage-clamp, fluorescence assay, 45Ca2+ isotopic uptake in Xenopus oocytes, site-directed mutagenesis, immunocytochemistry","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 1 — in vitro functional assay with mutagenesis in a heterologous expression system, single lab","pmids":["19955568"],"is_preprint":false},{"year":2012,"finding":"ZDHHC3 (DHHC3) is the enzyme responsible for palmitoylation of integrin α6 and β4 subunits; DHHC3 ablation markedly diminished integrin-dependent cellular cable formation on Matrigel, integrin signaling through Src, β4 phosphorylation at S1356 and S1424, and accelerated lysosomal degradation of α6β4 via increased cathepsin D exposure; rescued α6β4 accumulated intracellularly and could not reach the cell surface.","method":"RNAi knockdown, overexpression in multiple cell types, palmitoylation assay, Matrigel cable formation assay, phosphorylation analysis, lysosomal inhibitor (Pepstatin A) rescue","journal":"Cellular and molecular life sciences","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods across multiple cell types, clean KD with defined molecular and cellular phenotypes","pmids":["22314500"],"is_preprint":false},{"year":2012,"finding":"ZDHHC3 (GODZ) interacts with the death domain of TRAIL receptor DR4 (but not DR5) through its DHHC and C-terminal transmembrane domains, localizes DR4 to the plasma membrane, and regulates TRAIL/DR4-mediated apoptosis; mutation of the cysteine-rich motif of DR4 causes mistargeting and attenuates TRAIL-mediated apoptosis.","method":"SOS protein recruitment-yeast two-hybrid screening, pulldown, cell surface localization assay, apoptosis assay, DR4 cysteine mutagenesis","journal":"Cell death and differentiation","confidence":"Medium","confidence_rationale":"Tier 2-3 — Y2H identification plus functional rescue and mutagenesis, single lab","pmids":["22240897"],"is_preprint":false},{"year":2015,"finding":"ZDHHC3 undergoes autoacylation (palmitoylation) at the cysteine within the DHHC motif, as shown by mass spectrometry; conserved cysteines outside the DHHC motif coordinate two zinc ions per DHHC3 molecule, and chelation or mutation of these cysteines disrupts CRD structural integrity and causes enzymatic activity deficits.","method":"Mass spectrometry, site-directed mutagenesis of conserved CRD cysteines, limited proteolysis, metal chelation in vitro, fluorescent zinc indicator (mag-fura-2) stoichiometry measurement","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — in vitro biochemical reconstitution with mass spectrometry, mutagenesis, and structural perturbation analysis in a single study","pmids":["26487721"],"is_preprint":false},{"year":2016,"finding":"ZDHHC3 palmitoylates the neural cell adhesion molecule NCAM, and this activity is regulated by tyrosine phosphorylation: FGFR1 phosphorylates Tyr18 and Src phosphorylates Tyr295/Tyr297 of ZDHHC3; abrogation of these phosphorylation sites increased ZDHHC3 autopalmitoylation, enhanced interaction with NCAM, upregulated NCAM palmitoylation, and promoted neurite outgrowth in hippocampal neurons.","method":"Site-directed mutagenesis, pharmacological inhibition of FGFR and Src, cell-based and cell-free kinase assays, coimmunoprecipitation, neurite outgrowth assay in hippocampal neurons","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1-2 — mutagenesis of specific phosphorylation sites with both cell-based and cell-free validation, defined functional consequence in neurons","pmids":["27247265"],"is_preprint":false},{"year":2016,"finding":"In GODZ knockout mice, palmitoylation of γ2 GABA(A) receptor subunit and GAP-43 (growth-associated protein 43 kDa) was significantly reduced in brain; synaptic accumulation of GABA(A) receptors and GABAergic synaptic function were selectively reduced in GODZ KO neurons when competing with wild-type neurons; GODZ and SERZ-β localize to distinct cis- versus trans-Golgi compartments, explaining their different in vivo substrate specificities.","method":"Knockout mice (GODZ KO, SERZ-β KO, double KO), palmitoylation assay in brain, electrophysiology, immunocytochemistry, subcellular fractionation","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — clean in vivo KO with multiple orthogonal assays, mechanistic explanation for substrate specificity via Golgi compartment localization","pmids":["27875292"],"is_preprint":false},{"year":2016,"finding":"ZDHHC3 palmitoylates Gsα in Xenopus oocytes to maintain meiotic arrest at G2 phase; specific palmitoylation sites on Gsα were mapped, palmitoylation-deficient Gsα failed to maintain G2 arrest, and a critical ZDHHC3 residue required for its palmitoylation activity toward Gsα was identified; ZDHHC3 functions downstream of acsl1b in this pathway.","method":"Maternal RNA depletion in Xenopus oocytes, meiotic arrest assay, site-directed mutagenesis of Gsα palmitoylation sites and ZDHHC3 catalytic residue, progesterone threshold assay","journal":"Biology of reproduction","confidence":"Medium","confidence_rationale":"Tier 2 — genetic epistasis plus mutagenesis in Xenopus oocyte system with defined functional readout, single lab","pmids":["27512151"],"is_preprint":false},{"year":2017,"finding":"HSV-1 UL20 protein binds specifically to ZDHHC3 (GODZ) via two-hybrid and pulldown assays; ZDHHC3 palmitoylates UL20, and this palmitoylation is required for proper membrane targeting of UL20 and subsequent gK cell surface expression and HSV-1 infectivity; dominant-negative GODZ (C157S) and 2-bromopalmitate treatment reduced HSV-1 titers and altered UL20/gK localization.","method":"Yeast two-hybrid, pulldown assay, dominant-negative GODZ expression, palmitoylation assay, virus titer measurement, immunofluorescence","journal":"Journal of virology","confidence":"High","confidence_rationale":"Tier 2 — reciprocal binding assays plus functional palmitoylation assay plus dominant-negative and pharmacological inhibition with virus infectivity readout","pmids":["28724772"],"is_preprint":false},{"year":2017,"finding":"ZDHHC3 palmitoylates ERGIC3, and loss of DHHC3-dependent palmitoylation of ERGIC3 leads to upregulation of TXNIP, increased oxidative stress, and senescence in tumor cells; DHHC3 ablation reduced xenograft tumor growth and metastasis, and these effects required enzymatic activity (wild-type but not active-site-deficient DHHC3 reconstituted activity).","method":"ZDHHC3 ablation (RNAi/CRISPR) in MDA-MB-231 xenografts, gene array, fluorescence oxidative stress assay, reconstitution with WT vs. catalytic-dead ZDHHC3, concomitant TXNIP ablation","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 — in vivo xenograft plus enzymatic activity requirement demonstrated by catalytic-dead rescue, multiple orthogonal methods","pmids":["29055014"],"is_preprint":false},{"year":2018,"finding":"In GODZ−/− murine embryonic fibroblasts, HSV-1 replication is compromised: GODZ absence blocks palmitoylation of viral UL20, alters localization and expression of UL20, gK, gB, gC, and tegument/capsid proteins, and limits virion maturation at multiple steps as shown by electron microscopy; in vivo, ocularly infected GODZ−/− mice showed reduced corneal scarring and reduced HSV-1 latency reactivation.","method":"GODZ−/− knockout mouse-derived MEFs, palmitoylation assay, electron microscopy, immunofluorescence, in vivo ocular infection model","journal":"Journal of virology","confidence":"High","confidence_rationale":"Tier 2 — clean KO with multiple orthogonal readouts including ultrastructural analysis, in vitro and in vivo validation","pmids":["29187538"],"is_preprint":false},{"year":2020,"finding":"Mass spectrometry-based palmitoyl-proteomic analysis identified 22–28 antioxidant/redox-regulatory proteins as candidate ZDHHC3 substrates; DHHC3 ablation elevated oxidative stress and, combined with chemotherapeutic drug treatment, synergistically enhanced anti-growth effects, establishing DHHC3 as a regulator of antioxidant protein palmitoylation and cellular redox homeostasis.","method":"Comparative mass spectrometry-based palmitoyl-proteomics, DHHC3 ablation in breast and prostate cancer cell lines, fluorescence oxidative stress assays, cell proliferation assays","journal":"Cellular and molecular life sciences","confidence":"High","confidence_rationale":"Tier 1-2 — proteome-wide substrate identification by MS combined with functional phenotypic validation, multiple cell lines","pmids":["32986127"],"is_preprint":false},{"year":2021,"finding":"A high-throughput Acyl-cLIP enzymatic assay for ZDHHC3 (as well as ZDHHC7 and ZDHHC20) was developed and validated; in vitro Acyl-cLIP results were confirmed by cell-based palmitoylation assays, enabling inhibitor screening.","method":"Acylation-coupled lipophilic induction of polarization (Acyl-cLIP) in vitro assay, cell-based palmitoylation assay validation","journal":"ACS chemical biology","confidence":"Medium","confidence_rationale":"Tier 1 — in vitro enzymatic assay with cell-based validation, method development paper, single lab","pmids":["34374518"],"is_preprint":false},{"year":2022,"finding":"Structural exploration of human DHHC3 identified LAMTOR1 as an interacting protein by mass spectrometry and co-immunoprecipitation; cryo-EM imaging of the inactive hDHHS3 mutant showed a typical membrane protein side-view, providing initial structural characterization of DHHC3.","method":"Protein expression/purification, mass spectrometry, co-immunoprecipitation, cryo-EM","journal":"Polymers","confidence":"Low","confidence_rationale":"Tier 3 — preliminary structural data without functional validation, single Co-IP for LAMTOR1 interaction","pmids":["35893977"],"is_preprint":false},{"year":2023,"finding":"ZDHHC3 palmitoylates IRHOM2 at C476 within the iRhom homology domain, facilitating IRHOM2 cytomembrane translocation and stabilization; palmitoylation by ZDHHC3 (via its C157 DHHC domain) blocks TRIM31-mediated ubiquitin-proteasome degradation of IRHOM2; fatty acid challenge increases ZDHHC3–IRHOM2 association and IRHOM2 palmitoylation; hepatocyte-specific ZDHHC3 deletion suppresses NASH pathology in rodent and rabbit models.","method":"Co-immunoprecipitation, palmitoylation assay (acyl-RAC), site-directed mutagenesis (C476, C157), hepatocyte-specific KO mice, in vivo diet-induced NASH models, ubiquitination assay","journal":"Advanced science","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods including mutagenesis of both substrate and enzyme active site, in vitro and in vivo validation across two animal models","pmids":["37544908"],"is_preprint":false},{"year":2024,"finding":"ZDHHC3 palmitoylates PD-L1, maintaining its membrane stability; inhibition of ZDHHC3 enzymatic activity by benzosceptrin C (BC) prevents PD-L1 palmitoylation, causing PD-L1 to be transferred from the membrane to the cytoplasm where it cannot recycle via recycling endosomes and undergoes lysosome-mediated degradation.","method":"Cell-based palmitoylation assay, ZDHHC3 enzymatic inhibition by small molecule BC, flow cytometry and imaging of PD-L1 trafficking, T cell cytotoxicity assay, in vivo MC38 tumor model","journal":"Cell reports. Medicine","confidence":"High","confidence_rationale":"Tier 2 — mechanistic pathway dissected with enzymatic inhibitor plus trafficking assays plus in vivo validation","pmids":["38237597"],"is_preprint":false},{"year":2024,"finding":"ZDHHC3 S-acylates SCAP at C264, antagonizing HACE1-mediated SCAP ubiquitination and thereby promoting SCAP/SREBP2 signaling and cholesterol biosynthesis in HCC; SREBP2 transcriptionally upregulates ZDHHC3, forming a positive feedback loop; ABHD17A acts as the depalmitoylase counteracting ZDHHC3 on SCAP.","method":"Co-immunoprecipitation, palmitoylation assay, site-directed mutagenesis (C264), ubiquitination assay, chromatin immunoprecipitation (SREBP2), ZDHHC3 KO/overexpression, in vivo DEN/CCl4 HCC mouse model","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods with mutagenesis, feedback loop characterization, and in vivo validation","pmids":["39522165"],"is_preprint":false},{"year":2024,"finding":"ZDHHC3 palmitoylates PML/RARα oncofusion protein, and this palmitoylation regulates its oncogenic transcriptional activity and APL pathogenesis; knockdown or overexpression of ZDHHC3 respectively decreased or increased expression of proliferation- and differentiation-related genes; ZDHHC3 depletion or inhibition arrested malignant progression including in drug-resistant APL.","method":"Palmitoylation assay, ZDHHC3 knockdown/overexpression, gene expression analysis, APL cell differentiation/proliferation assays","journal":"Acta pharmacologica Sinica","confidence":"Medium","confidence_rationale":"Tier 2-3 — functional palmitoylation assay with KD/OE phenotype but limited mechanistic detail on palmitoylation site, single lab","pmids":["39227737"],"is_preprint":false},{"year":2024,"finding":"ZDHHC3 is required for palmitoylation of Cadm4 at C347, which stabilizes Cadm4 at the plasma membrane; genetic deletion of ZDHHC3 reduces Cadm4 palmitoylation and causes CNS myelination defects (loss, demyelination, hypermyelination) phenocopying C347A Cadm4 knock-in mice; altered Cadm4 palmitoylation impairs neuronal transmission, cognitive behaviors, and modulates WNT-β-Catenin–dependent oligodendrocyte differentiation.","method":"ZDHHC3 KO mice, Cadm4 C347A knock-in mice, palmitoylation assay, immunofluorescence, electrophysiology, behavioral tests, WNT-β-Catenin pathway analysis","journal":"Signal transduction and targeted therapy","confidence":"High","confidence_rationale":"Tier 2 — clean KO and KI mouse models with multiple orthogonal phenotypic readouts and pathway identification","pmids":["39327467"],"is_preprint":false},{"year":2024,"finding":"IL-1α signaling in precancerous epithelial dysplasia represses expression of DHHC3 (and DHHC7), enzymes responsible for palmitoylation of STING, thereby reducing STING-mediated type-I interferon signaling in myeloid cells.","method":"Genetically engineered mouse model, longitudinal human specimens, gene expression analysis, IL-1 blockade (pharmacological and genetic)","journal":"bioRxiv (preprint)","confidence":"Low","confidence_rationale":"Tier 3 — pathway placement by expression/genetic perturbation in complex in vivo model, STING palmitoylation by ZDHHC3 not directly demonstrated in this paper","pmids":[],"is_preprint":true},{"year":2026,"finding":"ZDHHC3 and ZDHHC7 mediate S-acylation of the small GTPase ARL15 at three conserved N-terminal cysteine residues (Cys17, Cys22, Cys23) in a partially redundant manner; loss of S-acylation disrupts ARL15 membrane association and redistributes it to the cytosol.","method":"APEGS assay, siRNA knockdown, CRISPR/Cas9 gene disruption, confocal imaging, subcellular fractionation, cysteine-to-serine mutagenesis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 — systematic stoichiometric palmitoylation assay, CRISPR KO, mutagenesis, and localization with functional consequence, multiple orthogonal methods","pmids":["41999893"],"is_preprint":false}],"current_model":"ZDHHC3 (GODZ) is a Golgi-localized DHHC-family palmitoyltransferase that catalyzes S-palmitoylation of diverse substrates—including GABA(A) receptor γ2 subunit, integrin α6β4, NCAM, DR4, PD-L1, SCAP, IRHOM2, Cadm4, Gsα, ARL15, and viral UL20—via a two-step autoacylation-then-transfer mechanism requiring the DHHC catalytic cysteine and zinc-coordinating CRD cysteines, with its activity further regulated by FGFR/Src-mediated tyrosine phosphorylation, thereby controlling substrate membrane localization, stability, trafficking, and downstream signaling in contexts ranging from GABAergic synaptogenesis and CNS myelination to tumor immune escape and viral infectivity."},"narrative":{"teleology":[{"year":2002,"claim":"Identification of ZDHHC3 as a Golgi-localized DHHC-domain protein that influences membrane protein trafficking established it as a candidate regulator of intracellular sorting.","evidence":"Overexpression in COS7 cells with immunofluorescence showing Golgi retention of GluRα1","pmids":["12163046"],"confidence":"Medium","gaps":["No enzymatic activity demonstrated","Overexpression artifact not excluded","Endogenous substrates unknown"]},{"year":2004,"claim":"Demonstrating that ZDHHC3 palmitoylates the GABA(A) receptor γ2 subunit via a specific cytoplasmic loop domain established ZDHHC3 as a bona fide palmitoyltransferase with a defined neuronal substrate.","evidence":"Yeast two-hybrid, metabolic palmitoylation assay, domain-dependency mutagenesis in heterologous cells and neurons","pmids":["15229235"],"confidence":"High","gaps":["In vivo relevance of GABA(A) receptor palmitoylation not yet shown","Substrate selectivity versus other DHHCs not addressed"]},{"year":2006,"claim":"Loss-of-function studies (dominant-negative C157S and RNAi) revealed that ZDHHC3 is selectively required for GABAergic synaptic receptor accumulation and inhibitory synaptic transmission, establishing its non-redundant role in synapse-type-specific function.","evidence":"Dominant-negative expression, RNAi, co-immunoprecipitation, electrophysiology in cultured neurons","pmids":["17151279"],"confidence":"High","gaps":["Knockout animal model not yet generated","Mechanism of selective GABAergic versus glutamatergic distinction unclear"]},{"year":2012,"claim":"Identification of integrin α6β4 and TRAIL receptor DR4 as ZDHHC3 substrates expanded its functional scope beyond neurons to cell adhesion, signaling, and apoptosis, revealing that palmitoylation by ZDHHC3 controls surface delivery and lysosomal degradation of diverse transmembrane proteins.","evidence":"RNAi knockdown across multiple cell types for integrin; Y2H, mutagenesis, and apoptosis assays for DR4","pmids":["22314500","22240897"],"confidence":"High","gaps":["DR4 palmitoylation site not directly mapped","In vivo validation of integrin and DR4 palmitoylation lacking"]},{"year":2015,"claim":"Biochemical reconstitution demonstrated that ZDHHC3 catalysis proceeds through autoacylation of the DHHC-motif cysteine, and that two zinc ions coordinated by conserved CRD cysteines are essential for structural integrity, establishing the enzymatic mechanism.","evidence":"Mass spectrometry detection of autoacylated intermediate, mutagenesis of CRD cysteines, metal chelation, zinc stoichiometry measurement","pmids":["26487721"],"confidence":"High","gaps":["No high-resolution atomic structure","Acyl-chain selectivity mechanism unknown","Substrate recognition determinants not structurally defined"]},{"year":2016,"claim":"Three concurrent advances established (1) FGFR1/Src-mediated tyrosine phosphorylation as a regulatory mechanism controlling ZDHHC3 autopalmitoylation and NCAM palmitoylation, (2) in vivo knockout validation that ZDHHC3 and ZDHHC7 occupy distinct Golgi compartments explaining substrate specificity, and (3) Gsα as a substrate linking ZDHHC3 to meiotic cell cycle arrest.","evidence":"Phospho-site mutagenesis and kinase assays for NCAM regulation; GODZ KO mice with brain palmitoyl-proteomics and electrophysiology; Xenopus oocyte maternal RNA depletion for Gsα","pmids":["27247265","27875292","27512151"],"confidence":"High","gaps":["Phosphorylation regulation not validated in vivo","Cis- vs trans-Golgi specificity structural basis unknown","Gsα pathway relevance in mammalian systems not demonstrated"]},{"year":2017,"claim":"ZDHHC3 was shown to palmitoylate HSV-1 UL20 protein, with loss of this palmitoylation disrupting viral glycoprotein surface expression and reducing viral infectivity, establishing ZDHHC3 as a host factor exploited by herpesviruses.","evidence":"Y2H, pulldown, dominant-negative GODZ, palmitoylation assay, virus titer measurement; subsequently confirmed in GODZ−/− MEFs and ocular infection model","pmids":["28724772","29187538"],"confidence":"High","gaps":["Whether other DHHCs compensate in vivo not fully addressed","Mechanism of viral UL20 recognition by ZDHHC3 unknown"]},{"year":2017,"claim":"Identification of ERGIC3 palmitoylation by ZDHHC3 linked its enzymatic activity to cellular redox homeostasis: loss of ZDHHC3 upregulated TXNIP, elevated oxidative stress, and suppressed xenograft tumor growth, positioning ZDHHC3 as a cancer-relevant palmitoylation enzyme.","evidence":"ZDHHC3 ablation (RNAi/CRISPR) in xenografts, catalytic-dead reconstitution, gene array, oxidative stress assay","pmids":["29055014"],"confidence":"High","gaps":["Direct palmitoylation site on ERGIC3 not mapped","Mechanism linking ERGIC3 palmitoylation to TXNIP expression unclear"]},{"year":2020,"claim":"Proteome-wide palmitoyl-proteomics identified 22–28 antioxidant/redox proteins as ZDHHC3 substrates, generalizing its role in redox homeostasis and demonstrating synergy between ZDHHC3 loss and chemotherapy.","evidence":"Comparative MS-based palmitoyl-proteomics in breast and prostate cancer cell lines with DHHC3 ablation","pmids":["32986127"],"confidence":"High","gaps":["Most candidate substrates not individually validated","In vivo therapeutic relevance of combination strategy not tested"]},{"year":2023,"claim":"ZDHHC3 palmitoylation of IRHOM2 at C476 was shown to block TRIM31-mediated ubiquitin-proteasome degradation, with hepatocyte-specific ZDHHC3 deletion suppressing diet-induced NASH in rodents, linking ZDHHC3 to metabolic liver disease.","evidence":"Palmitoylation assay (acyl-RAC), mutagenesis of both enzyme and substrate, hepatocyte-specific KO mice, NASH diet models in mice and rabbits","pmids":["37544908"],"confidence":"High","gaps":["Human relevance of ZDHHC3–IRHOM2 axis in NASH not demonstrated","Downstream IRHOM2 effectors mediating hepatic pathology incompletely defined"]},{"year":2024,"claim":"Multiple studies converged to define ZDHHC3 as a regulator of immune checkpoint (PD-L1), cholesterol metabolism (SCAP/SREBP2 feedback loop), CNS myelination (Cadm4/WNT-β-Catenin), and leukemia (PML/RARα), demonstrating that palmitoylation-dependent stabilization against lysosomal or proteasomal degradation is a unifying mechanism across substrates.","evidence":"PD-L1: enzymatic inhibitor BC with trafficking/degradation assays and tumor model; SCAP: mutagenesis of C264, ubiquitination assay, ChIP for SREBP2, HCC mouse model; Cadm4: ZDHHC3 KO and C347A KI mice with electrophysiology and behavioral tests; PML/RARα: KD/OE with differentiation assays","pmids":["38237597","39522165","39327467","39227737"],"confidence":"High","gaps":["PML/RARα palmitoylation site not mapped","Relative contribution of ZDHHC3 vs ZDHHC7 to PD-L1 palmitoylation in vivo unclear","SCAP feedback loop not validated in human HCC tissue"]},{"year":2026,"claim":"ZDHHC3 was shown to S-acylate the small GTPase ARL15 at three N-terminal cysteines in a partially redundant manner with ZDHHC7, with loss of acylation redistributing ARL15 from membranes to the cytosol.","evidence":"APEGS assay, CRISPR KO, siRNA, cysteine mutagenesis, confocal imaging, subcellular fractionation","pmids":["41999893"],"confidence":"High","gaps":["Functional consequence of ARL15 mislocalization not fully characterized","Redundancy with ZDHHC7 makes ZDHHC3-specific contribution hard to isolate in vivo"]},{"year":null,"claim":"A high-resolution atomic structure of ZDHHC3 in complex with a substrate or acyl-CoA is still lacking, and the structural determinants of its broad but selective substrate recognition remain undefined.","evidence":"","pmids":[],"confidence":"High","gaps":["No atomic-resolution structure available","Substrate recognition code not defined","Tissue-specific regulation of ZDHHC3 expression and activity incompletely understood"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[1,4,6,7,8,9,10,11,13,16,17,18,20,22]}],"localization":[{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[0,1,8]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[1,6,7,13]},{"term_id":"R-HSA-112316","term_label":"Neuronal System","supporting_discovery_ids":[1,2,8,20]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[7,17,18,20]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[17]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[18]}],"complexes":[],"partners":["GABRG2","NCAM1","ITGB4","IRHOM2","SCAP","CD274","CADM4","ARL15"],"other_free_text":[]},"mechanistic_narrative":"ZDHHC3 (GODZ) is a Golgi-resident DHHC-family protein S-acyltransferase (palmitoyltransferase) that catalyzes the transfer of palmitoyl groups to a broad spectrum of substrates, thereby controlling their membrane localization, stability, trafficking, and downstream signaling across neural, immune, metabolic, and viral contexts. The enzyme operates via a two-step mechanism in which the catalytic DHHC-motif cysteine (C157) first undergoes autoacylation before transferring the acyl chain to substrate cysteines, with two zinc ions coordinated by conserved cysteine-rich domain residues being essential for structural integrity and activity [PMID:26487721]. In the CNS, ZDHHC3 palmitoylates the GABA(A) receptor γ2 subunit to promote its synaptic accumulation and GABAergic inhibitory transmission [PMID:15229235, PMID:27875292], palmitoylates NCAM under regulation by FGFR1/Src-mediated tyrosine phosphorylation to modulate neurite outgrowth [PMID:27247265], and palmitoylates Cadm4 at C347 to stabilize it at the plasma membrane and support CNS myelination and oligodendrocyte differentiation via WNT-β-Catenin signaling [PMID:39327467]. Beyond the nervous system, ZDHHC3 palmitoylates diverse substrates including integrin α6β4 to control surface trafficking and Src signaling [PMID:22314500], PD-L1 to maintain its plasma membrane stability and enable tumor immune evasion [PMID:38237597], SCAP at C264 to antagonize ubiquitination and promote SREBP2-dependent cholesterol biosynthesis in a transcriptional positive feedback loop [PMID:39522165], IRHOM2 to block TRIM31-mediated degradation and promote NASH pathogenesis [PMID:37544908], and HSV-1 UL20 to facilitate viral glycoprotein trafficking and infectivity [PMID:28724772, PMID:29187538]."},"prefetch_data":{"uniprot":{"accession":"Q9NYG2","full_name":"Palmitoyltransferase ZDHHC3","aliases":["Acyltransferase ZDHHC3","Protein DHHC1","Zinc finger DHHC domain-containing protein 3","DHHC-3"],"length_aa":299,"mass_kda":34.2,"function":"Golgi-localized palmitoyltransferase that catalyzes the addition of palmitate onto various protein substrates (PubMed:19001095, PubMed:21926431, PubMed:22240897, PubMed:22314500, PubMed:23034182). Has no stringent fatty acid selectivity and in addition to palmitate can also transfer onto target proteins myristate from tetradecanoyl-CoA and stearate from octadecanoyl-CoA (By similarity). Plays an important role in G protein-coupled receptor signaling pathways involving GNAQ and potentially other heterotrimeric G proteins by regulating their dynamic association with the plasma membrane (PubMed:19001095). Palmitoylates ITGA6 and ITGB4, thereby regulating the alpha-6/beta-4 integrin localization, expression and function in cell adhesion to laminin (PubMed:22314500). Plays a role in the TRAIL-activated apoptotic signaling pathway most probably through the palmitoylation and localization to the plasma membrane of TNFRSF10A (PubMed:22240897). In the brain, by palmitoylating the gamma subunit GABRG2 of GABA(A) receptors and regulating their postsynaptic accumulation, plays a role in synaptic GABAergic inhibitory function and GABAergic innervation (By similarity). Palmitoylates the neuronal protein GAP43 which is also involved in the formation of GABAergic synapses (By similarity). Palmitoylates NCDN thereby regulating its association with endosome membranes (By similarity). Probably palmitoylates PRCD and is involved in its proper localization within the photoreceptor (By similarity). Could mediate the palmitoylation of NCAM1 and regulate neurite outgrowth (By similarity). Could palmitoylate DNAJC5 and regulate its localization to Golgi membranes (By similarity). Also constitutively palmitoylates DLG4 (By similarity). May also palmitoylate SNAP25 (By similarity). Could palmitoylate the glutamate receptors GRIA1 and GRIA2 but this has not been confirmed in vivo (By similarity). Could also palmitoylate the D(2) dopamine receptor DRD2 (PubMed:26535572). May also palmitoylate LAMTOR1, promoting its localization to lysosomal membranes (PubMed:35893977). Palmitoylates the Toll-like receptor 9/TLR9 in the Golgi and thereby regulates TLR9 trafficking to endosomes (PubMed:38169466). May palmitoylate CALHM1 and CALHM3 subunits of gustatory voltage-gated ion channels and modulate channel gating and kinetics May also function as a calcium transporter","subcellular_location":"Golgi apparatus membrane","url":"https://www.uniprot.org/uniprotkb/Q9NYG2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ZDHHC3","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"COPB2","stoichiometry":0.2},{"gene":"GORASP2","stoichiometry":0.2},{"gene":"TMED10","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/ZDHHC3","total_profiled":1310},"omim":[{"mim_id":"621547","title":"ZDHHC PALMITOYLTRANSFERASE 4; ZDHHC4","url":"https://www.omim.org/entry/621547"},{"mim_id":"617334","title":"ZDHHC PALMITOYLTRANSFERASE 23; ZDHHC23","url":"https://www.omim.org/entry/617334"},{"mim_id":"617150","title":"ZDHHC PALMITOYLTRANSFERASE 3; ZDHHC3","url":"https://www.omim.org/entry/617150"},{"mim_id":"608784","title":"ZDHHC PALMITOYLTRANSFERASE 8; ZDHHC8","url":"https://www.omim.org/entry/608784"},{"mim_id":"190090","title":"SRC PROTOONCOGENE, NONRECEPTOR TYROSINE KINASE; SRC","url":"https://www.omim.org/entry/190090"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Golgi apparatus","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ZDHHC3"},"hgnc":{"alias_symbol":["ZNF373","GODZ","DHHC3"],"prev_symbol":[]},"alphafold":{"accession":"Q9NYG2","domains":[{"cath_id":"1.20.140","chopping":"46-103_167-280","consensus_level":"high","plddt":94.3001,"start":46,"end":280}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NYG2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NYG2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NYG2-F1-predicted_aligned_error_v6.png","plddt_mean":85.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ZDHHC3","jax_strain_url":"https://www.jax.org/strain/search?query=ZDHHC3"},"sequence":{"accession":"Q9NYG2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NYG2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NYG2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NYG2"}},"corpus_meta":[{"pmid":"15229235","id":"PMC_15229235","title":"The gamma2 subunit of GABA(A) receptors is a substrate for palmitoylation by GODZ.","date":"2004","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/15229235","citation_count":203,"is_preprint":false},{"pmid":"17151279","id":"PMC_17151279","title":"GODZ-mediated palmitoylation of GABA(A) receptors is required for normal assembly and function of GABAergic inhibitory synapses.","date":"2006","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/17151279","citation_count":137,"is_preprint":false},{"pmid":"29055014","id":"PMC_29055014","title":"Protein Acyltransferase DHHC3 Regulates Breast Tumor Growth, Oxidative Stress, and Senescence.","date":"2017","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/29055014","citation_count":67,"is_preprint":false},{"pmid":"38237597","id":"PMC_38237597","title":"Benzosceptrin C induces lysosomal degradation of PD-L1 and promotes antitumor immunity by targeting DHHC3.","date":"2024","source":"Cell reports. 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1 UL20 to GODZ (DHHC3) Affects Its Palmitoylation and Is Essential for Infectivity and Proper Targeting and Localization of UL20 and Glycoprotein K.","date":"2017","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/28724772","citation_count":18,"is_preprint":false},{"pmid":"32986127","id":"PMC_32986127","title":"Antioxidant functions of DHHC3 suppress anti-cancer drug activities.","date":"2020","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/32986127","citation_count":17,"is_preprint":false},{"pmid":"39349454","id":"PMC_39349454","title":"Treating ICB-resistant cancer by inhibiting PD-L1 via DHHC3 degradation induced by cell penetrating peptide-induced chimera conjugates.","date":"2024","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/39349454","citation_count":16,"is_preprint":false},{"pmid":"39327467","id":"PMC_39327467","title":"Palmitoylation regulates myelination by modulating the ZDHHC3-Cadm4 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mediated by the Golgi S-acyltransferases ZDHHC7 and ZDHHC3.","date":"2026","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/41999893","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.12.06.626475","title":"SOX2-induced IL1α-mediated immune suppression drives epithelial dysplasia malignant transformation","date":"2024-12-10","source":"bioRxiv","url":"https://doi.org/10.1101/2024.12.06.626475","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":17112,"output_tokens":6328,"usd":0.073128},"stage2":{"model":"claude-opus-4-6","input_tokens":10001,"output_tokens":3593,"usd":0.209745},"total_usd":0.282873,"stage1_batch_id":"msgbatch_01BparvgRZ9S2LMidvaoo86K","stage2_batch_id":"msgbatch_01NkBfv9SVZWthhS7j6mU6G3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2002,\n      \"finding\": \"GODZ (ZDHHC3) is a Golgi apparatus-specific protein with a DHHC zinc finger domain and four putative transmembrane regions; overexpression in COS7 cells suppressed sorting of glutamate receptor GluRα1 from the Golgi apparatus, implicating ZDHHC3 in membrane protein trafficking.\",\n      \"method\": \"Overexpression in COS7 cells, subcellular localization by immunofluorescence\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single overexpression experiment with defined trafficking phenotype, single lab\",\n      \"pmids\": [\"12163046\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"ZDHHC3 (GODZ) was identified as a palmitoyltransferase that palmitoylates the γ2 subunit of GABA(A) receptors via interaction with a conserved 14-amino acid cysteine-rich domain in the large cytoplasmic loop of γ1-3 subunits; GODZ localizes to the Golgi complex in neurons and its coexpression with GABA(A) receptors in heterologous cells results in γ2 palmitoylation in a cytoplasmic loop domain-dependent manner.\",\n      \"method\": \"SOS-recruitment yeast two-hybrid, coexpression in heterologous cells, metabolic palmitoylation assay, immunofluorescence localization\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (Y2H, palmitoylation assay, domain-dependency mutagenesis), replicated in neurons, highly cited\",\n      \"pmids\": [\"15229235\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"ZDHHC3 (GODZ) and its paralog SERZ-β (ZDHHC7) form homomultimers and heteromultimers; GODZ is required for normal synaptic accumulation of GABA(A) receptors, whole-cell and synaptic GABAergic inhibitory function, and GABAergic innervation, but is dispensable for postsynaptic AMPA receptor-mediated glutamatergic transmission; dominant-negative GODZ (C157S) and GODZ-specific RNAi established these requirements.\",\n      \"method\": \"Coimmunoprecipitation, in vivo cross-linking, dominant-negative expression (GODZ C157S), plasmid-based RNAi, electrophysiology in neurons\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP plus clean KD with defined synaptic phenotype, multiple orthogonal methods, replicated\",\n      \"pmids\": [\"17151279\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"ZDHHC3 (GODZ) mediates Ca2+ transport in Xenopus oocytes in a palmitoylation-dependent manner; mutation of the DHHC motif (GODZ-DHHS) or treatment with palmitoylation inhibitor 2-bromopalmitate reduced Ca2+ transport ~80%; a separate V61R mutation abolished Ca2+ transport without affecting palmitoyl acyltransferase activity, indicating dual functions.\",\n      \"method\": \"Two-electrode voltage-clamp, fluorescence assay, 45Ca2+ isotopic uptake in Xenopus oocytes, site-directed mutagenesis, immunocytochemistry\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 — in vitro functional assay with mutagenesis in a heterologous expression system, single lab\",\n      \"pmids\": [\"19955568\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"ZDHHC3 (DHHC3) is the enzyme responsible for palmitoylation of integrin α6 and β4 subunits; DHHC3 ablation markedly diminished integrin-dependent cellular cable formation on Matrigel, integrin signaling through Src, β4 phosphorylation at S1356 and S1424, and accelerated lysosomal degradation of α6β4 via increased cathepsin D exposure; rescued α6β4 accumulated intracellularly and could not reach the cell surface.\",\n      \"method\": \"RNAi knockdown, overexpression in multiple cell types, palmitoylation assay, Matrigel cable formation assay, phosphorylation analysis, lysosomal inhibitor (Pepstatin A) rescue\",\n      \"journal\": \"Cellular and molecular life sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods across multiple cell types, clean KD with defined molecular and cellular phenotypes\",\n      \"pmids\": [\"22314500\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"ZDHHC3 (GODZ) interacts with the death domain of TRAIL receptor DR4 (but not DR5) through its DHHC and C-terminal transmembrane domains, localizes DR4 to the plasma membrane, and regulates TRAIL/DR4-mediated apoptosis; mutation of the cysteine-rich motif of DR4 causes mistargeting and attenuates TRAIL-mediated apoptosis.\",\n      \"method\": \"SOS protein recruitment-yeast two-hybrid screening, pulldown, cell surface localization assay, apoptosis assay, DR4 cysteine mutagenesis\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — Y2H identification plus functional rescue and mutagenesis, single lab\",\n      \"pmids\": [\"22240897\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"ZDHHC3 undergoes autoacylation (palmitoylation) at the cysteine within the DHHC motif, as shown by mass spectrometry; conserved cysteines outside the DHHC motif coordinate two zinc ions per DHHC3 molecule, and chelation or mutation of these cysteines disrupts CRD structural integrity and causes enzymatic activity deficits.\",\n      \"method\": \"Mass spectrometry, site-directed mutagenesis of conserved CRD cysteines, limited proteolysis, metal chelation in vitro, fluorescent zinc indicator (mag-fura-2) stoichiometry measurement\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro biochemical reconstitution with mass spectrometry, mutagenesis, and structural perturbation analysis in a single study\",\n      \"pmids\": [\"26487721\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"ZDHHC3 palmitoylates the neural cell adhesion molecule NCAM, and this activity is regulated by tyrosine phosphorylation: FGFR1 phosphorylates Tyr18 and Src phosphorylates Tyr295/Tyr297 of ZDHHC3; abrogation of these phosphorylation sites increased ZDHHC3 autopalmitoylation, enhanced interaction with NCAM, upregulated NCAM palmitoylation, and promoted neurite outgrowth in hippocampal neurons.\",\n      \"method\": \"Site-directed mutagenesis, pharmacological inhibition of FGFR and Src, cell-based and cell-free kinase assays, coimmunoprecipitation, neurite outgrowth assay in hippocampal neurons\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — mutagenesis of specific phosphorylation sites with both cell-based and cell-free validation, defined functional consequence in neurons\",\n      \"pmids\": [\"27247265\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"In GODZ knockout mice, palmitoylation of γ2 GABA(A) receptor subunit and GAP-43 (growth-associated protein 43 kDa) was significantly reduced in brain; synaptic accumulation of GABA(A) receptors and GABAergic synaptic function were selectively reduced in GODZ KO neurons when competing with wild-type neurons; GODZ and SERZ-β localize to distinct cis- versus trans-Golgi compartments, explaining their different in vivo substrate specificities.\",\n      \"method\": \"Knockout mice (GODZ KO, SERZ-β KO, double KO), palmitoylation assay in brain, electrophysiology, immunocytochemistry, subcellular fractionation\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean in vivo KO with multiple orthogonal assays, mechanistic explanation for substrate specificity via Golgi compartment localization\",\n      \"pmids\": [\"27875292\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"ZDHHC3 palmitoylates Gsα in Xenopus oocytes to maintain meiotic arrest at G2 phase; specific palmitoylation sites on Gsα were mapped, palmitoylation-deficient Gsα failed to maintain G2 arrest, and a critical ZDHHC3 residue required for its palmitoylation activity toward Gsα was identified; ZDHHC3 functions downstream of acsl1b in this pathway.\",\n      \"method\": \"Maternal RNA depletion in Xenopus oocytes, meiotic arrest assay, site-directed mutagenesis of Gsα palmitoylation sites and ZDHHC3 catalytic residue, progesterone threshold assay\",\n      \"journal\": \"Biology of reproduction\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis plus mutagenesis in Xenopus oocyte system with defined functional readout, single lab\",\n      \"pmids\": [\"27512151\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"HSV-1 UL20 protein binds specifically to ZDHHC3 (GODZ) via two-hybrid and pulldown assays; ZDHHC3 palmitoylates UL20, and this palmitoylation is required for proper membrane targeting of UL20 and subsequent gK cell surface expression and HSV-1 infectivity; dominant-negative GODZ (C157S) and 2-bromopalmitate treatment reduced HSV-1 titers and altered UL20/gK localization.\",\n      \"method\": \"Yeast two-hybrid, pulldown assay, dominant-negative GODZ expression, palmitoylation assay, virus titer measurement, immunofluorescence\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal binding assays plus functional palmitoylation assay plus dominant-negative and pharmacological inhibition with virus infectivity readout\",\n      \"pmids\": [\"28724772\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"ZDHHC3 palmitoylates ERGIC3, and loss of DHHC3-dependent palmitoylation of ERGIC3 leads to upregulation of TXNIP, increased oxidative stress, and senescence in tumor cells; DHHC3 ablation reduced xenograft tumor growth and metastasis, and these effects required enzymatic activity (wild-type but not active-site-deficient DHHC3 reconstituted activity).\",\n      \"method\": \"ZDHHC3 ablation (RNAi/CRISPR) in MDA-MB-231 xenografts, gene array, fluorescence oxidative stress assay, reconstitution with WT vs. catalytic-dead ZDHHC3, concomitant TXNIP ablation\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo xenograft plus enzymatic activity requirement demonstrated by catalytic-dead rescue, multiple orthogonal methods\",\n      \"pmids\": [\"29055014\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"In GODZ−/− murine embryonic fibroblasts, HSV-1 replication is compromised: GODZ absence blocks palmitoylation of viral UL20, alters localization and expression of UL20, gK, gB, gC, and tegument/capsid proteins, and limits virion maturation at multiple steps as shown by electron microscopy; in vivo, ocularly infected GODZ−/− mice showed reduced corneal scarring and reduced HSV-1 latency reactivation.\",\n      \"method\": \"GODZ−/− knockout mouse-derived MEFs, palmitoylation assay, electron microscopy, immunofluorescence, in vivo ocular infection model\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with multiple orthogonal readouts including ultrastructural analysis, in vitro and in vivo validation\",\n      \"pmids\": [\"29187538\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Mass spectrometry-based palmitoyl-proteomic analysis identified 22–28 antioxidant/redox-regulatory proteins as candidate ZDHHC3 substrates; DHHC3 ablation elevated oxidative stress and, combined with chemotherapeutic drug treatment, synergistically enhanced anti-growth effects, establishing DHHC3 as a regulator of antioxidant protein palmitoylation and cellular redox homeostasis.\",\n      \"method\": \"Comparative mass spectrometry-based palmitoyl-proteomics, DHHC3 ablation in breast and prostate cancer cell lines, fluorescence oxidative stress assays, cell proliferation assays\",\n      \"journal\": \"Cellular and molecular life sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — proteome-wide substrate identification by MS combined with functional phenotypic validation, multiple cell lines\",\n      \"pmids\": [\"32986127\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"A high-throughput Acyl-cLIP enzymatic assay for ZDHHC3 (as well as ZDHHC7 and ZDHHC20) was developed and validated; in vitro Acyl-cLIP results were confirmed by cell-based palmitoylation assays, enabling inhibitor screening.\",\n      \"method\": \"Acylation-coupled lipophilic induction of polarization (Acyl-cLIP) in vitro assay, cell-based palmitoylation assay validation\",\n      \"journal\": \"ACS chemical biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 — in vitro enzymatic assay with cell-based validation, method development paper, single lab\",\n      \"pmids\": [\"34374518\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Structural exploration of human DHHC3 identified LAMTOR1 as an interacting protein by mass spectrometry and co-immunoprecipitation; cryo-EM imaging of the inactive hDHHS3 mutant showed a typical membrane protein side-view, providing initial structural characterization of DHHC3.\",\n      \"method\": \"Protein expression/purification, mass spectrometry, co-immunoprecipitation, cryo-EM\",\n      \"journal\": \"Polymers\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — preliminary structural data without functional validation, single Co-IP for LAMTOR1 interaction\",\n      \"pmids\": [\"35893977\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ZDHHC3 palmitoylates IRHOM2 at C476 within the iRhom homology domain, facilitating IRHOM2 cytomembrane translocation and stabilization; palmitoylation by ZDHHC3 (via its C157 DHHC domain) blocks TRIM31-mediated ubiquitin-proteasome degradation of IRHOM2; fatty acid challenge increases ZDHHC3–IRHOM2 association and IRHOM2 palmitoylation; hepatocyte-specific ZDHHC3 deletion suppresses NASH pathology in rodent and rabbit models.\",\n      \"method\": \"Co-immunoprecipitation, palmitoylation assay (acyl-RAC), site-directed mutagenesis (C476, C157), hepatocyte-specific KO mice, in vivo diet-induced NASH models, ubiquitination assay\",\n      \"journal\": \"Advanced science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods including mutagenesis of both substrate and enzyme active site, in vitro and in vivo validation across two animal models\",\n      \"pmids\": [\"37544908\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZDHHC3 palmitoylates PD-L1, maintaining its membrane stability; inhibition of ZDHHC3 enzymatic activity by benzosceptrin C (BC) prevents PD-L1 palmitoylation, causing PD-L1 to be transferred from the membrane to the cytoplasm where it cannot recycle via recycling endosomes and undergoes lysosome-mediated degradation.\",\n      \"method\": \"Cell-based palmitoylation assay, ZDHHC3 enzymatic inhibition by small molecule BC, flow cytometry and imaging of PD-L1 trafficking, T cell cytotoxicity assay, in vivo MC38 tumor model\",\n      \"journal\": \"Cell reports. Medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic pathway dissected with enzymatic inhibitor plus trafficking assays plus in vivo validation\",\n      \"pmids\": [\"38237597\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZDHHC3 S-acylates SCAP at C264, antagonizing HACE1-mediated SCAP ubiquitination and thereby promoting SCAP/SREBP2 signaling and cholesterol biosynthesis in HCC; SREBP2 transcriptionally upregulates ZDHHC3, forming a positive feedback loop; ABHD17A acts as the depalmitoylase counteracting ZDHHC3 on SCAP.\",\n      \"method\": \"Co-immunoprecipitation, palmitoylation assay, site-directed mutagenesis (C264), ubiquitination assay, chromatin immunoprecipitation (SREBP2), ZDHHC3 KO/overexpression, in vivo DEN/CCl4 HCC mouse model\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods with mutagenesis, feedback loop characterization, and in vivo validation\",\n      \"pmids\": [\"39522165\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZDHHC3 palmitoylates PML/RARα oncofusion protein, and this palmitoylation regulates its oncogenic transcriptional activity and APL pathogenesis; knockdown or overexpression of ZDHHC3 respectively decreased or increased expression of proliferation- and differentiation-related genes; ZDHHC3 depletion or inhibition arrested malignant progression including in drug-resistant APL.\",\n      \"method\": \"Palmitoylation assay, ZDHHC3 knockdown/overexpression, gene expression analysis, APL cell differentiation/proliferation assays\",\n      \"journal\": \"Acta pharmacologica Sinica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — functional palmitoylation assay with KD/OE phenotype but limited mechanistic detail on palmitoylation site, single lab\",\n      \"pmids\": [\"39227737\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZDHHC3 is required for palmitoylation of Cadm4 at C347, which stabilizes Cadm4 at the plasma membrane; genetic deletion of ZDHHC3 reduces Cadm4 palmitoylation and causes CNS myelination defects (loss, demyelination, hypermyelination) phenocopying C347A Cadm4 knock-in mice; altered Cadm4 palmitoylation impairs neuronal transmission, cognitive behaviors, and modulates WNT-β-Catenin–dependent oligodendrocyte differentiation.\",\n      \"method\": \"ZDHHC3 KO mice, Cadm4 C347A knock-in mice, palmitoylation assay, immunofluorescence, electrophysiology, behavioral tests, WNT-β-Catenin pathway analysis\",\n      \"journal\": \"Signal transduction and targeted therapy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean KO and KI mouse models with multiple orthogonal phenotypic readouts and pathway identification\",\n      \"pmids\": [\"39327467\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"IL-1α signaling in precancerous epithelial dysplasia represses expression of DHHC3 (and DHHC7), enzymes responsible for palmitoylation of STING, thereby reducing STING-mediated type-I interferon signaling in myeloid cells.\",\n      \"method\": \"Genetically engineered mouse model, longitudinal human specimens, gene expression analysis, IL-1 blockade (pharmacological and genetic)\",\n      \"journal\": \"bioRxiv (preprint)\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — pathway placement by expression/genetic perturbation in complex in vivo model, STING palmitoylation by ZDHHC3 not directly demonstrated in this paper\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"ZDHHC3 and ZDHHC7 mediate S-acylation of the small GTPase ARL15 at three conserved N-terminal cysteine residues (Cys17, Cys22, Cys23) in a partially redundant manner; loss of S-acylation disrupts ARL15 membrane association and redistributes it to the cytosol.\",\n      \"method\": \"APEGS assay, siRNA knockdown, CRISPR/Cas9 gene disruption, confocal imaging, subcellular fractionation, cysteine-to-serine mutagenesis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — systematic stoichiometric palmitoylation assay, CRISPR KO, mutagenesis, and localization with functional consequence, multiple orthogonal methods\",\n      \"pmids\": [\"41999893\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ZDHHC3 (GODZ) is a Golgi-localized DHHC-family palmitoyltransferase that catalyzes S-palmitoylation of diverse substrates—including GABA(A) receptor γ2 subunit, integrin α6β4, NCAM, DR4, PD-L1, SCAP, IRHOM2, Cadm4, Gsα, ARL15, and viral UL20—via a two-step autoacylation-then-transfer mechanism requiring the DHHC catalytic cysteine and zinc-coordinating CRD cysteines, with its activity further regulated by FGFR/Src-mediated tyrosine phosphorylation, thereby controlling substrate membrane localization, stability, trafficking, and downstream signaling in contexts ranging from GABAergic synaptogenesis and CNS myelination to tumor immune escape and viral infectivity.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"ZDHHC3 (GODZ) is a Golgi-resident DHHC-family protein S-acyltransferase (palmitoyltransferase) that catalyzes the transfer of palmitoyl groups to a broad spectrum of substrates, thereby controlling their membrane localization, stability, trafficking, and downstream signaling across neural, immune, metabolic, and viral contexts. The enzyme operates via a two-step mechanism in which the catalytic DHHC-motif cysteine (C157) first undergoes autoacylation before transferring the acyl chain to substrate cysteines, with two zinc ions coordinated by conserved cysteine-rich domain residues being essential for structural integrity and activity [PMID:26487721]. In the CNS, ZDHHC3 palmitoylates the GABA(A) receptor γ2 subunit to promote its synaptic accumulation and GABAergic inhibitory transmission [PMID:15229235, PMID:27875292], palmitoylates NCAM under regulation by FGFR1/Src-mediated tyrosine phosphorylation to modulate neurite outgrowth [PMID:27247265], and palmitoylates Cadm4 at C347 to stabilize it at the plasma membrane and support CNS myelination and oligodendrocyte differentiation via WNT-β-Catenin signaling [PMID:39327467]. Beyond the nervous system, ZDHHC3 palmitoylates diverse substrates including integrin α6β4 to control surface trafficking and Src signaling [PMID:22314500], PD-L1 to maintain its plasma membrane stability and enable tumor immune evasion [PMID:38237597], SCAP at C264 to antagonize ubiquitination and promote SREBP2-dependent cholesterol biosynthesis in a transcriptional positive feedback loop [PMID:39522165], IRHOM2 to block TRIM31-mediated degradation and promote NASH pathogenesis [PMID:37544908], and HSV-1 UL20 to facilitate viral glycoprotein trafficking and infectivity [PMID:28724772, PMID:29187538].\",\n  \"teleology\": [\n    {\n      \"year\": 2002,\n      \"claim\": \"Identification of ZDHHC3 as a Golgi-localized DHHC-domain protein that influences membrane protein trafficking established it as a candidate regulator of intracellular sorting.\",\n      \"evidence\": \"Overexpression in COS7 cells with immunofluorescence showing Golgi retention of GluRα1\",\n      \"pmids\": [\"12163046\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No enzymatic activity demonstrated\", \"Overexpression artifact not excluded\", \"Endogenous substrates unknown\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Demonstrating that ZDHHC3 palmitoylates the GABA(A) receptor γ2 subunit via a specific cytoplasmic loop domain established ZDHHC3 as a bona fide palmitoyltransferase with a defined neuronal substrate.\",\n      \"evidence\": \"Yeast two-hybrid, metabolic palmitoylation assay, domain-dependency mutagenesis in heterologous cells and neurons\",\n      \"pmids\": [\"15229235\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance of GABA(A) receptor palmitoylation not yet shown\", \"Substrate selectivity versus other DHHCs not addressed\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Loss-of-function studies (dominant-negative C157S and RNAi) revealed that ZDHHC3 is selectively required for GABAergic synaptic receptor accumulation and inhibitory synaptic transmission, establishing its non-redundant role in synapse-type-specific function.\",\n      \"evidence\": \"Dominant-negative expression, RNAi, co-immunoprecipitation, electrophysiology in cultured neurons\",\n      \"pmids\": [\"17151279\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Knockout animal model not yet generated\", \"Mechanism of selective GABAergic versus glutamatergic distinction unclear\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Identification of integrin α6β4 and TRAIL receptor DR4 as ZDHHC3 substrates expanded its functional scope beyond neurons to cell adhesion, signaling, and apoptosis, revealing that palmitoylation by ZDHHC3 controls surface delivery and lysosomal degradation of diverse transmembrane proteins.\",\n      \"evidence\": \"RNAi knockdown across multiple cell types for integrin; Y2H, mutagenesis, and apoptosis assays for DR4\",\n      \"pmids\": [\"22314500\", \"22240897\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"DR4 palmitoylation site not directly mapped\", \"In vivo validation of integrin and DR4 palmitoylation lacking\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Biochemical reconstitution demonstrated that ZDHHC3 catalysis proceeds through autoacylation of the DHHC-motif cysteine, and that two zinc ions coordinated by conserved CRD cysteines are essential for structural integrity, establishing the enzymatic mechanism.\",\n      \"evidence\": \"Mass spectrometry detection of autoacylated intermediate, mutagenesis of CRD cysteines, metal chelation, zinc stoichiometry measurement\",\n      \"pmids\": [\"26487721\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No high-resolution atomic structure\", \"Acyl-chain selectivity mechanism unknown\", \"Substrate recognition determinants not structurally defined\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Three concurrent advances established (1) FGFR1/Src-mediated tyrosine phosphorylation as a regulatory mechanism controlling ZDHHC3 autopalmitoylation and NCAM palmitoylation, (2) in vivo knockout validation that ZDHHC3 and ZDHHC7 occupy distinct Golgi compartments explaining substrate specificity, and (3) Gsα as a substrate linking ZDHHC3 to meiotic cell cycle arrest.\",\n      \"evidence\": \"Phospho-site mutagenesis and kinase assays for NCAM regulation; GODZ KO mice with brain palmitoyl-proteomics and electrophysiology; Xenopus oocyte maternal RNA depletion for Gsα\",\n      \"pmids\": [\"27247265\", \"27875292\", \"27512151\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Phosphorylation regulation not validated in vivo\", \"Cis- vs trans-Golgi specificity structural basis unknown\", \"Gsα pathway relevance in mammalian systems not demonstrated\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"ZDHHC3 was shown to palmitoylate HSV-1 UL20 protein, with loss of this palmitoylation disrupting viral glycoprotein surface expression and reducing viral infectivity, establishing ZDHHC3 as a host factor exploited by herpesviruses.\",\n      \"evidence\": \"Y2H, pulldown, dominant-negative GODZ, palmitoylation assay, virus titer measurement; subsequently confirmed in GODZ−/− MEFs and ocular infection model\",\n      \"pmids\": [\"28724772\", \"29187538\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether other DHHCs compensate in vivo not fully addressed\", \"Mechanism of viral UL20 recognition by ZDHHC3 unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identification of ERGIC3 palmitoylation by ZDHHC3 linked its enzymatic activity to cellular redox homeostasis: loss of ZDHHC3 upregulated TXNIP, elevated oxidative stress, and suppressed xenograft tumor growth, positioning ZDHHC3 as a cancer-relevant palmitoylation enzyme.\",\n      \"evidence\": \"ZDHHC3 ablation (RNAi/CRISPR) in xenografts, catalytic-dead reconstitution, gene array, oxidative stress assay\",\n      \"pmids\": [\"29055014\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct palmitoylation site on ERGIC3 not mapped\", \"Mechanism linking ERGIC3 palmitoylation to TXNIP expression unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Proteome-wide palmitoyl-proteomics identified 22–28 antioxidant/redox proteins as ZDHHC3 substrates, generalizing its role in redox homeostasis and demonstrating synergy between ZDHHC3 loss and chemotherapy.\",\n      \"evidence\": \"Comparative MS-based palmitoyl-proteomics in breast and prostate cancer cell lines with DHHC3 ablation\",\n      \"pmids\": [\"32986127\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Most candidate substrates not individually validated\", \"In vivo therapeutic relevance of combination strategy not tested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"ZDHHC3 palmitoylation of IRHOM2 at C476 was shown to block TRIM31-mediated ubiquitin-proteasome degradation, with hepatocyte-specific ZDHHC3 deletion suppressing diet-induced NASH in rodents, linking ZDHHC3 to metabolic liver disease.\",\n      \"evidence\": \"Palmitoylation assay (acyl-RAC), mutagenesis of both enzyme and substrate, hepatocyte-specific KO mice, NASH diet models in mice and rabbits\",\n      \"pmids\": [\"37544908\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Human relevance of ZDHHC3–IRHOM2 axis in NASH not demonstrated\", \"Downstream IRHOM2 effectors mediating hepatic pathology incompletely defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Multiple studies converged to define ZDHHC3 as a regulator of immune checkpoint (PD-L1), cholesterol metabolism (SCAP/SREBP2 feedback loop), CNS myelination (Cadm4/WNT-β-Catenin), and leukemia (PML/RARα), demonstrating that palmitoylation-dependent stabilization against lysosomal or proteasomal degradation is a unifying mechanism across substrates.\",\n      \"evidence\": \"PD-L1: enzymatic inhibitor BC with trafficking/degradation assays and tumor model; SCAP: mutagenesis of C264, ubiquitination assay, ChIP for SREBP2, HCC mouse model; Cadm4: ZDHHC3 KO and C347A KI mice with electrophysiology and behavioral tests; PML/RARα: KD/OE with differentiation assays\",\n      \"pmids\": [\"38237597\", \"39522165\", \"39327467\", \"39227737\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"PML/RARα palmitoylation site not mapped\", \"Relative contribution of ZDHHC3 vs ZDHHC7 to PD-L1 palmitoylation in vivo unclear\", \"SCAP feedback loop not validated in human HCC tissue\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"ZDHHC3 was shown to S-acylate the small GTPase ARL15 at three N-terminal cysteines in a partially redundant manner with ZDHHC7, with loss of acylation redistributing ARL15 from membranes to the cytosol.\",\n      \"evidence\": \"APEGS assay, CRISPR KO, siRNA, cysteine mutagenesis, confocal imaging, subcellular fractionation\",\n      \"pmids\": [\"41999893\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of ARL15 mislocalization not fully characterized\", \"Redundancy with ZDHHC7 makes ZDHHC3-specific contribution hard to isolate in vivo\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A high-resolution atomic structure of ZDHHC3 in complex with a substrate or acyl-CoA is still lacking, and the structural determinants of its broad but selective substrate recognition remain undefined.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No atomic-resolution structure available\", \"Substrate recognition code not defined\", \"Tissue-specific regulation of ZDHHC3 expression and activity incompletely understood\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [1, 4, 6, 7, 8, 9, 10, 11, 13, 16, 17, 18, 20, 22]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [0, 1, 8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": []},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [1, 6, 7, 13]},\n      {\"term_id\": \"R-HSA-112316\", \"supporting_discovery_ids\": [1, 2, 8, 20]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [7, 17, 18, 20]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [17]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [18]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"GABRG2\",\n      \"NCAM1\",\n      \"ITGB4\",\n      \"IRHOM2\",\n      \"SCAP\",\n      \"CD274\",\n      \"CADM4\",\n      \"ARL15\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}