{"gene":"PEX10","run_date":"2026-06-10T05:19:53","timeline":{"discoveries":[{"year":1998,"finding":"PEX10 encodes an integral peroxisomal membrane protein with two transmembrane segments and a C3HC4 zinc-finger RING motif; both N- and C-terminal regions are exposed to the cytosol. Its expression rescues peroxisomal matrix-protein import in PBD complementation group B/CG7 fibroblasts, and the C-terminal RING finger domain is required for biological function (frameshift removing RING motif abolished rescue activity).","method":"Epitope-tagged expression constructs, immunofluorescence, subcellular fractionation, functional complementation assay in patient fibroblasts, mutant cDNA rescue experiments","journal":"Human molecular genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct functional complementation plus topology determination by epitope-tagging, replicated across two independent groups (PMID:9700193 and PMID:9683594)","pmids":["9700193","9683594"],"is_preprint":false},{"year":1999,"finding":"PEX10 physically interacts with PEX12 (via PEX12's zinc-RING domain) and this interaction is biologically relevant: overexpression of PEX10 suppresses a PEX12 missense mutation (S320F). Loss of PEX10 does not prevent PEX5 docking to peroxisomes, placing PEX10 downstream of receptor docking in peroxisomal matrix-protein import.","method":"Two-hybrid assay, blot overlay assay, co-immunoprecipitation, genetic suppressor overexpression, PEX5 peroxisome-association assay","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal binding confirmed by three orthogonal methods (Y2H, blot overlay, Co-IP) plus in vivo genetic suppression establishing biological relevance","pmids":["10562279"],"is_preprint":false},{"year":2000,"finding":"The C-terminal zinc-binding (RING) domain of PEX10 is critical for PEX10 function; missense mutation H290Q in this domain results in a milder (residual) PEX10 activity compared with null alleles. Nonsense/frameshift mutations deleting the C-terminal two-thirds still yielded unexpectedly high activity in complementation assays, attributed to read-through or alternative translation, but deliberate deletion or mutation of downstream sequence abolished this activity.","method":"Functional complementation assay in PEX10-deficient patient fibroblasts, site-directed mutagenesis of PEX10 cDNA constructs","journal":"Human mutation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional complementation with multiple mutant constructs, single lab, but results complicated by assay artifacts noted by the authors","pmids":["10862081"],"is_preprint":false},{"year":2010,"finding":"In Arabidopsis (plant ortholog study), abolishing Zn2+ coordination in PEX10's RING finger (ΔZn mutation) causes deformed peroxisomal shapes, impaired peroxisome-chloroplast contact, and defective photorespiration, revealing that the N-terminal cytosolic domain of PEX10 is critical for peroxisome biogenesis and shape, while the RING finger domain is implicated in ubiquitination of PEX5.","method":"Dominant-negative overexpression of zinc-binding mutants in Arabidopsis, confocal microscopy of peroxisome morphology, metabolite analysis, photorespiration assay under varying CO2","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal readouts (morphology, metabolomics, gas exchange) in plant ortholog; plant context limits direct extrapolation to mammalian PEX10","pmids":["20679226"],"is_preprint":false},{"year":2024,"finding":"In prostate cancer cells, PEX10 expression is induced by ROS activators and reduces intracellular ROS levels. The androgen receptor (AR), in cooperation with FOXA1, acts as a transcriptional enhancer to promote PEX10 expression. Enzalutamide inhibits AR function and thereby suppresses PEX10, sensitizing cells to ROS-activating agents (ML210, RSL3).","method":"ROS measurement assays, ChIP/enhancer analysis for AR-FOXA1 at PEX10 locus, enzalutamide treatment, knockdown/overexpression of PEX10, cell viability assays with ROS activators","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct transcriptional regulation demonstrated by enhancer/ChIP and functional ROS assays; single lab, cancer-cell context","pmids":["39097593"],"is_preprint":false},{"year":2025,"finding":"Structural and functional evidence (preprint) indicates that the Pex2/Pex10/Pex12 complex functions as an E3-ubiquitin ligase complex at the peroxisomal membrane that assembles with Pex5/Pex8 to initiate recycling of the PTS1 receptor Pex5 following cargo translocation.","method":"Cryo-EM structure of Pex8-Pex5 complex, functional complementation assays in yeast, mutagenesis of interaction interfaces","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — structural determination with functional validation, but preprint and PEX10 role is inferred from complex membership rather than directly interrogated","pmids":[],"is_preprint":true}],"current_model":"PEX10 is an integral peroxisomal membrane protein with two transmembrane segments and a cytosol-facing C3HC4 zinc-finger RING domain that functions as part of the Pex2/Pex10/Pex12 E3-ubiquitin ligase complex; it physically interacts with PEX12 and acts downstream of PEX5 receptor docking to mediate peroxisomal matrix-protein import and PEX5 receptor recycling, with its RING domain essential for function and its expression transcriptionally regulated by the androgen receptor/FOXA1 axis."},"narrative":{"mechanistic_narrative":"PEX10 is an integral peroxisomal membrane protein required for peroxisomal matrix-protein import, defined by two transmembrane segments and a cytosol-facing C3HC4 zinc-finger RING motif whose integrity is essential for function [PMID:9700193, PMID:9683594]. It operates downstream of PEX5 receptor docking: loss of PEX10 does not prevent PEX5 association with peroxisomes, and PEX10 instead physically interacts with PEX12 through PEX12's zinc-RING domain, with PEX10 overexpression suppressing a PEX12 missense allele—establishing PEX10 and PEX12 as functionally coupled RING partners [PMID:10562279]. The RING domain is the critical catalytic element: zinc-coordination mutants impair function, and abolishing Zn2+ binding in the plant ortholog produces deformed peroxisomes, defective peroxisome–chloroplast contact, and failed photorespiration, linking the RING to ubiquitination of PEX5 [PMID:10862081, PMID:20679226]. In prostate cancer cells PEX10 is transcriptionally induced through an androgen-receptor/FOXA1 enhancer and lowers intracellular ROS, so that AR blockade by enzalutamide suppresses PEX10 and sensitizes cells to ROS-activating agents [PMID:39097593]. Beyond these findings, the detailed enzymatic mechanism of PEX10-mediated ubiquitin transfer in mammalian cells has not been directly characterized in the available corpus.","teleology":[{"year":1998,"claim":"Established that PEX10 is a peroxisomal membrane protein whose RING domain is required to support matrix-protein import, defining it as a biogenesis factor rather than an imported matrix protein.","evidence":"Epitope-tagged topology mapping, subcellular fractionation, and functional complementation of PBD group B/CG7 fibroblasts with wild-type and RING-deleted cDNAs","pmids":["9700193","9683594"],"confidence":"High","gaps":["Molecular activity of the RING domain not yet defined","Direct binding partners at the membrane not identified"]},{"year":1999,"claim":"Placed PEX10 in a defined import step by showing it acts downstream of PEX5 docking and physically partners with PEX12, answering where in the import cascade PEX10 functions.","evidence":"Two-hybrid, blot overlay, co-immunoprecipitation, in vivo genetic suppression of a PEX12 mutation, and PEX5 peroxisome-association assay","pmids":["10562279"],"confidence":"High","gaps":["Biochemical consequence of the PEX10–PEX12 interaction not resolved","Whether the pair acts catalytically not addressed here"]},{"year":2000,"claim":"Refined which residues of the RING domain carry function by showing graded activity loss across missense versus null alleles, connecting genotype to residual biological activity.","evidence":"Site-directed mutagenesis and functional complementation in PEX10-deficient patient fibroblasts","pmids":["10862081"],"confidence":"Medium","gaps":["Complementation results confounded by read-through/alternative-translation artifacts noted by authors","Single-lab assay without orthogonal biochemical confirmation"]},{"year":2010,"claim":"Linked the RING domain to PEX5 ubiquitination and revealed the N-terminal cytosolic domain controls peroxisome shape and inter-organelle contact, broadening PEX10's role beyond import to organelle morphology and metabolism.","evidence":"Dominant-negative zinc-binding mutants in Arabidopsis with confocal morphology, metabolite analysis, and photorespiration gas-exchange assays","pmids":["20679226"],"confidence":"Medium","gaps":["Plant ortholog context limits direct extrapolation to mammalian PEX10","Direct demonstration of PEX10-catalyzed PEX5 ubiquitination not shown"]},{"year":2024,"claim":"Identified a transcriptional input and a physiological output for PEX10, showing AR/FOXA1 drives its expression and PEX10 buffers intracellular ROS, connecting peroxisomal function to redox control in cancer cells.","evidence":"ChIP/enhancer analysis of AR-FOXA1 at the PEX10 locus, ROS assays, enzalutamide treatment, and PEX10 knockdown/overexpression with viability readouts","pmids":["39097593"],"confidence":"Medium","gaps":["Mechanism linking PEX10 to ROS reduction not defined","Single cancer-cell context"]},{"year":2025,"claim":"Provided structural framing for the Pex2/Pex10/Pex12 RING complex acting as an E3 ligase that assembles with Pex5/Pex8 to initiate PTS1-receptor recycling after cargo translocation.","evidence":"Cryo-EM of Pex8–Pex5 complex with yeast complementation and interface mutagenesis (preprint)","pmids":[],"confidence":"Medium","gaps":["PEX10's specific role inferred from complex membership rather than directly interrogated","Preprint, not peer-reviewed","Mammalian relevance not established"]},{"year":null,"claim":"The direct enzymatic activity of mammalian PEX10 as a ubiquitin ligase—its E2 partner, substrate-engagement mechanism, and the structure of the assembled Pex2/Pex10/Pex12 complex—remains uncharacterized in the available corpus.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No reconstituted ubiquitination assay with mammalian PEX10","Cognate E2 enzyme not identified","No mammalian structure of the assembled RING complex"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[0,1,3]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[3]}],"localization":[],"pathway":[{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[0,1]},{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[3]}],"complexes":["Pex2/Pex10/Pex12 E3-ubiquitin ligase complex"],"partners":["PEX12","PEX5"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O60683","full_name":"Peroxisome biogenesis factor 10","aliases":["Peroxin-10","Peroxisomal biogenesis factor 10","Peroxisome assembly protein 10","RING finger protein 69"],"length_aa":326,"mass_kda":37.1,"function":"E3 ubiquitin-protein ligase component of a retrotranslocation channel required for peroxisome organization by mediating export of the PEX5 receptor from peroxisomes to the cytosol, thereby promoting PEX5 recycling (PubMed:24662292). The retrotranslocation channel is composed of PEX2, PEX10 and PEX12; each subunit contributing transmembrane segments that coassemble into an open channel that specifically allows the passage of PEX5 through the peroxisomal membrane (By similarity). PEX10 also regulates PEX5 recycling by acting as a E3 ubiquitin-protein ligase (PubMed:24662292). When PEX5 recycling is compromised, PEX10 catalyzes polyubiquitination of PEX5 during its passage through the retrotranslocation channel, leading to its degradation (By similarity)","subcellular_location":"Peroxisome membrane","url":"https://www.uniprot.org/uniprotkb/O60683/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/PEX10","classification":"Not Classified","n_dependent_lines":75,"n_total_lines":1208,"dependency_fraction":0.062086092715231786},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/PEX10","total_profiled":1310},"omim":[{"mim_id":"614885","title":"PEROXISOME BIOGENESIS DISORDER 11B; PBD11B","url":"https://www.omim.org/entry/614885"},{"mim_id":"614871","title":"PEROXISOME BIOGENESIS DISORDER 6B; PBD6B","url":"https://www.omim.org/entry/614871"},{"mim_id":"614870","title":"PEROXISOME BIOGENESIS DISORDER 6A (ZELLWEGER); PBD6A","url":"https://www.omim.org/entry/614870"},{"mim_id":"602859","title":"PEROXISOME BIOGENESIS FACTOR 10; PEX10","url":"https://www.omim.org/entry/602859"},{"mim_id":"601789","title":"PEROXISOME BIOGENESIS FACTOR 13; PEX13","url":"https://www.omim.org/entry/601789"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/PEX10"},"hgnc":{"alias_symbol":["RNF69"],"prev_symbol":[]},"alphafold":{"accession":"O60683","domains":[{"cath_id":"-","chopping":"5-121_145-241","consensus_level":"high","plddt":88.1304,"start":5,"end":241},{"cath_id":"3.30.40.10","chopping":"271-326","consensus_level":"medium","plddt":87.7102,"start":271,"end":326}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O60683","model_url":"https://alphafold.ebi.ac.uk/files/AF-O60683-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O60683-F1-predicted_aligned_error_v6.png","plddt_mean":82.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=PEX10","jax_strain_url":"https://www.jax.org/strain/search?query=PEX10"},"sequence":{"accession":"O60683","fasta_url":"https://rest.uniprot.org/uniprotkb/O60683.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O60683/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O60683"}},"corpus_meta":[{"pmid":"10562279","id":"PMC_10562279","title":"PEX12 interacts with PEX5 and PEX10 and acts downstream of receptor docking in peroxisomal matrix protein import.","date":"1999","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/10562279","citation_count":117,"is_preprint":false},{"pmid":"9683594","id":"PMC_9683594","title":"Identification of PEX10, the gene defective in complementation group 7 of the peroxisome-biogenesis disorders.","date":"1998","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/9683594","citation_count":103,"is_preprint":false},{"pmid":"9700193","id":"PMC_9700193","title":"Mutations in PEX10 is the cause of Zellweger peroxisome deficiency syndrome of complementation group B.","date":"1998","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/9700193","citation_count":89,"is_preprint":false},{"pmid":"20679226","id":"PMC_20679226","title":"Different functions of the C3HC4 zinc RING finger peroxins PEX10, PEX2, and PEX12 in peroxisome formation and matrix protein import.","date":"2010","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/20679226","citation_count":48,"is_preprint":false},{"pmid":"17041890","id":"PMC_17041890","title":"Identification of novel mutations in PEX2, PEX6, PEX10, PEX12, and PEX13 in Zellweger spectrum patients.","date":"2006","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/17041890","citation_count":39,"is_preprint":false},{"pmid":"16388862","id":"PMC_16388862","title":"Identification and characterization of three peroxins--PEX6, PEX10 and PEX12--involved in glycosome biogenesis in Trypanosoma brucei.","date":"2005","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/16388862","citation_count":37,"is_preprint":false},{"pmid":"34132586","id":"PMC_34132586","title":"Increased Accumulation of Squalene in Engineered Yarrowia lipolytica through Deletion of PEX10 and URE2.","date":"2021","source":"Applied and environmental microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/34132586","citation_count":32,"is_preprint":false},{"pmid":"19127411","id":"PMC_19127411","title":"A PEX10 defect in a patient with no detectable defect in peroxisome assembly or metabolism in cultured fibroblasts.","date":"2008","source":"Journal of inherited metabolic disease","url":"https://pubmed.ncbi.nlm.nih.gov/19127411","citation_count":31,"is_preprint":false},{"pmid":"12794690","id":"PMC_12794690","title":"Genetic heterogeneity of peroxisome biogenesis disorders among Japanese patients: evidence for a founder haplotype for the most common PEX10 gene mutation.","date":"2003","source":"American journal of medical genetics. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/12794690","citation_count":25,"is_preprint":false},{"pmid":"10862081","id":"PMC_10862081","title":"Phenotype-genotype relationships in PEX10-deficient peroxisome biogenesis disorder patients.","date":"2000","source":"Human mutation","url":"https://pubmed.ncbi.nlm.nih.gov/10862081","citation_count":22,"is_preprint":false},{"pmid":"24648396","id":"PMC_24648396","title":"Association study between polymorphisms of PRMT6, PEX10, SOX5, and nonobstructive azoospermia in the Han Chinese population.","date":"2014","source":"Biology of reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/24648396","citation_count":19,"is_preprint":false},{"pmid":"30863997","id":"PMC_30863997","title":"PEX10, SIRPA-SIRPG, and SOX5 gene polymorphisms are strongly associated with nonobstructive azoospermia susceptibility.","date":"2019","source":"Journal of assisted reproduction and genetics","url":"https://pubmed.ncbi.nlm.nih.gov/30863997","citation_count":17,"is_preprint":false},{"pmid":"28320181","id":"PMC_28320181","title":"Ataxic form of autosomal recessive PEX10-related peroxisome biogenesis disorders with a novel compound heterozygous gene mutation and characteristic clinical phenotype.","date":"2017","source":"Journal of the neurological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/28320181","citation_count":9,"is_preprint":false},{"pmid":"28784167","id":"PMC_28784167","title":"Identification of a novel mutation in PEX10 in a patient with attenuated Zellweger spectrum disorder: a case report.","date":"2017","source":"Journal of medical case reports","url":"https://pubmed.ncbi.nlm.nih.gov/28784167","citation_count":8,"is_preprint":false},{"pmid":"39097593","id":"PMC_39097593","title":"Enzalutamide inhibits PEX10 function and sensitizes prostate cancer cells to ROS activators.","date":"2024","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/39097593","citation_count":6,"is_preprint":false},{"pmid":"30640048","id":"PMC_30640048","title":"Ataxia with novel compound heterozygous PEX10 mutations and a literature review of PEX10-related peroxisome biogenesis disorders.","date":"2019","source":"Clinical neurology and neurosurgery","url":"https://pubmed.ncbi.nlm.nih.gov/30640048","citation_count":6,"is_preprint":false},{"pmid":"35038753","id":"PMC_35038753","title":"How to Detect Isolated PEX10-Related Cerebellar Ataxia?","date":"2022","source":"Neuropediatrics","url":"https://pubmed.ncbi.nlm.nih.gov/35038753","citation_count":3,"is_preprint":false},{"pmid":"40267090","id":"PMC_40267090","title":"Identification of novel compound heterozygous variants in the PEX10 gene in a Han-Chinese family with PEX10-related peroxisome biogenesis disorders.","date":"2025","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/40267090","citation_count":3,"is_preprint":false},{"pmid":"25636090","id":"PMC_25636090","title":"[Abnormal expression of PEX10 gene may be related to epilepsy associated with 1p36 copy number variations].","date":"2015","source":"Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/25636090","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.08.30.673231","title":"Structure of Pex8 in complex with peroxisomal receptor Pex5 reveals its essential role in peroxisomal cargo translocation","date":"2025-09-02","source":"bioRxiv","url":"https://doi.org/10.1101/2025.08.30.673231","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.05.18.654701","title":"Engineering precision zebrafish alleles of human disease","date":"2025-05-21","source":"bioRxiv","url":"https://doi.org/10.1101/2025.05.18.654701","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":12586,"output_tokens":1762,"usd":0.032094,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8559,"output_tokens":2449,"usd":0.05201,"stage2_stop_reason":"end_turn"},"total_usd":0.084104,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1998,\n      \"finding\": \"PEX10 encodes an integral peroxisomal membrane protein with two transmembrane segments and a C3HC4 zinc-finger RING motif; both N- and C-terminal regions are exposed to the cytosol. Its expression rescues peroxisomal matrix-protein import in PBD complementation group B/CG7 fibroblasts, and the C-terminal RING finger domain is required for biological function (frameshift removing RING motif abolished rescue activity).\",\n      \"method\": \"Epitope-tagged expression constructs, immunofluorescence, subcellular fractionation, functional complementation assay in patient fibroblasts, mutant cDNA rescue experiments\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct functional complementation plus topology determination by epitope-tagging, replicated across two independent groups (PMID:9700193 and PMID:9683594)\",\n      \"pmids\": [\"9700193\", \"9683594\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"PEX10 physically interacts with PEX12 (via PEX12's zinc-RING domain) and this interaction is biologically relevant: overexpression of PEX10 suppresses a PEX12 missense mutation (S320F). Loss of PEX10 does not prevent PEX5 docking to peroxisomes, placing PEX10 downstream of receptor docking in peroxisomal matrix-protein import.\",\n      \"method\": \"Two-hybrid assay, blot overlay assay, co-immunoprecipitation, genetic suppressor overexpression, PEX5 peroxisome-association assay\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal binding confirmed by three orthogonal methods (Y2H, blot overlay, Co-IP) plus in vivo genetic suppression establishing biological relevance\",\n      \"pmids\": [\"10562279\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"The C-terminal zinc-binding (RING) domain of PEX10 is critical for PEX10 function; missense mutation H290Q in this domain results in a milder (residual) PEX10 activity compared with null alleles. Nonsense/frameshift mutations deleting the C-terminal two-thirds still yielded unexpectedly high activity in complementation assays, attributed to read-through or alternative translation, but deliberate deletion or mutation of downstream sequence abolished this activity.\",\n      \"method\": \"Functional complementation assay in PEX10-deficient patient fibroblasts, site-directed mutagenesis of PEX10 cDNA constructs\",\n      \"journal\": \"Human mutation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional complementation with multiple mutant constructs, single lab, but results complicated by assay artifacts noted by the authors\",\n      \"pmids\": [\"10862081\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"In Arabidopsis (plant ortholog study), abolishing Zn2+ coordination in PEX10's RING finger (ΔZn mutation) causes deformed peroxisomal shapes, impaired peroxisome-chloroplast contact, and defective photorespiration, revealing that the N-terminal cytosolic domain of PEX10 is critical for peroxisome biogenesis and shape, while the RING finger domain is implicated in ubiquitination of PEX5.\",\n      \"method\": \"Dominant-negative overexpression of zinc-binding mutants in Arabidopsis, confocal microscopy of peroxisome morphology, metabolite analysis, photorespiration assay under varying CO2\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal readouts (morphology, metabolomics, gas exchange) in plant ortholog; plant context limits direct extrapolation to mammalian PEX10\",\n      \"pmids\": [\"20679226\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In prostate cancer cells, PEX10 expression is induced by ROS activators and reduces intracellular ROS levels. The androgen receptor (AR), in cooperation with FOXA1, acts as a transcriptional enhancer to promote PEX10 expression. Enzalutamide inhibits AR function and thereby suppresses PEX10, sensitizing cells to ROS-activating agents (ML210, RSL3).\",\n      \"method\": \"ROS measurement assays, ChIP/enhancer analysis for AR-FOXA1 at PEX10 locus, enzalutamide treatment, knockdown/overexpression of PEX10, cell viability assays with ROS activators\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct transcriptional regulation demonstrated by enhancer/ChIP and functional ROS assays; single lab, cancer-cell context\",\n      \"pmids\": [\"39097593\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Structural and functional evidence (preprint) indicates that the Pex2/Pex10/Pex12 complex functions as an E3-ubiquitin ligase complex at the peroxisomal membrane that assembles with Pex5/Pex8 to initiate recycling of the PTS1 receptor Pex5 following cargo translocation.\",\n      \"method\": \"Cryo-EM structure of Pex8-Pex5 complex, functional complementation assays in yeast, mutagenesis of interaction interfaces\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — structural determination with functional validation, but preprint and PEX10 role is inferred from complex membership rather than directly interrogated\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"PEX10 is an integral peroxisomal membrane protein with two transmembrane segments and a cytosol-facing C3HC4 zinc-finger RING domain that functions as part of the Pex2/Pex10/Pex12 E3-ubiquitin ligase complex; it physically interacts with PEX12 and acts downstream of PEX5 receptor docking to mediate peroxisomal matrix-protein import and PEX5 receptor recycling, with its RING domain essential for function and its expression transcriptionally regulated by the androgen receptor/FOXA1 axis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"PEX10 is an integral peroxisomal membrane protein required for peroxisomal matrix-protein import, defined by two transmembrane segments and a cytosol-facing C3HC4 zinc-finger RING motif whose integrity is essential for function [#0]. It operates downstream of PEX5 receptor docking: loss of PEX10 does not prevent PEX5 association with peroxisomes, and PEX10 instead physically interacts with PEX12 through PEX12's zinc-RING domain, with PEX10 overexpression suppressing a PEX12 missense allele—establishing PEX10 and PEX12 as functionally coupled RING partners [#1]. The RING domain is the critical catalytic element: zinc-coordination mutants impair function, and abolishing Zn2+ binding in the plant ortholog produces deformed peroxisomes, defective peroxisome–chloroplast contact, and failed photorespiration, linking the RING to ubiquitination of PEX5 [#2, #3]. In prostate cancer cells PEX10 is transcriptionally induced through an androgen-receptor/FOXA1 enhancer and lowers intracellular ROS, so that AR blockade by enzalutamide suppresses PEX10 and sensitizes cells to ROS-activating agents [#4]. Beyond these findings, the detailed enzymatic mechanism of PEX10-mediated ubiquitin transfer in mammalian cells has not been directly characterized in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Established that PEX10 is a peroxisomal membrane protein whose RING domain is required to support matrix-protein import, defining it as a biogenesis factor rather than an imported matrix protein.\",\n      \"evidence\": \"Epitope-tagged topology mapping, subcellular fractionation, and functional complementation of PBD group B/CG7 fibroblasts with wild-type and RING-deleted cDNAs\",\n      \"pmids\": [\"9700193\", \"9683594\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular activity of the RING domain not yet defined\", \"Direct binding partners at the membrane not identified\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Placed PEX10 in a defined import step by showing it acts downstream of PEX5 docking and physically partners with PEX12, answering where in the import cascade PEX10 functions.\",\n      \"evidence\": \"Two-hybrid, blot overlay, co-immunoprecipitation, in vivo genetic suppression of a PEX12 mutation, and PEX5 peroxisome-association assay\",\n      \"pmids\": [\"10562279\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Biochemical consequence of the PEX10–PEX12 interaction not resolved\", \"Whether the pair acts catalytically not addressed here\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Refined which residues of the RING domain carry function by showing graded activity loss across missense versus null alleles, connecting genotype to residual biological activity.\",\n      \"evidence\": \"Site-directed mutagenesis and functional complementation in PEX10-deficient patient fibroblasts\",\n      \"pmids\": [\"10862081\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Complementation results confounded by read-through/alternative-translation artifacts noted by authors\", \"Single-lab assay without orthogonal biochemical confirmation\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Linked the RING domain to PEX5 ubiquitination and revealed the N-terminal cytosolic domain controls peroxisome shape and inter-organelle contact, broadening PEX10's role beyond import to organelle morphology and metabolism.\",\n      \"evidence\": \"Dominant-negative zinc-binding mutants in Arabidopsis with confocal morphology, metabolite analysis, and photorespiration gas-exchange assays\",\n      \"pmids\": [\"20679226\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Plant ortholog context limits direct extrapolation to mammalian PEX10\", \"Direct demonstration of PEX10-catalyzed PEX5 ubiquitination not shown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified a transcriptional input and a physiological output for PEX10, showing AR/FOXA1 drives its expression and PEX10 buffers intracellular ROS, connecting peroxisomal function to redox control in cancer cells.\",\n      \"evidence\": \"ChIP/enhancer analysis of AR-FOXA1 at the PEX10 locus, ROS assays, enzalutamide treatment, and PEX10 knockdown/overexpression with viability readouts\",\n      \"pmids\": [\"39097593\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking PEX10 to ROS reduction not defined\", \"Single cancer-cell context\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Provided structural framing for the Pex2/Pex10/Pex12 RING complex acting as an E3 ligase that assembles with Pex5/Pex8 to initiate PTS1-receptor recycling after cargo translocation.\",\n      \"evidence\": \"Cryo-EM of Pex8–Pex5 complex with yeast complementation and interface mutagenesis (preprint)\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"PEX10's specific role inferred from complex membership rather than directly interrogated\", \"Preprint, not peer-reviewed\", \"Mammalian relevance not established\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The direct enzymatic activity of mammalian PEX10 as a ubiquitin ligase—its E2 partner, substrate-engagement mechanism, and the structure of the assembled Pex2/Pex10/Pex12 complex—remains uncharacterized in the available corpus.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No reconstituted ubiquitination assay with mammalian PEX10\", \"Cognate E2 enzyme not identified\", \"No mammalian structure of the assembled RING complex\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [0, 1, 3]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005778\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"complexes\": [\"Pex2/Pex10/Pex12 E3-ubiquitin ligase complex\"],\n    \"partners\": [\"PEX12\", \"PEX5\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":4,"faith_pct":100.0}}