{"gene":"PEX10","run_date":"2026-04-29T11:37:58","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. Loss of PEX10 function blocks peroxisomal matrix protein import while peroxisome membrane protein import and peroxisome formation remain intact.","method":"Expression of epitope-tagged PEX10 in patient fibroblasts, functional complementation assay, mutation analysis","journal":"Human molecular genetics","confidence":"High","confidence_rationale":"Tier 1–2 — topology mapping by expression of epitope-tagged protein, functional complementation with defined mutations, replicated across two independent labs (PMID:9700193 and PMID:9683594)","pmids":["9700193","9683594"],"is_preprint":false},{"year":1999,"finding":"PEX12 physically interacts with both PEX5 (PTS1 receptor) and PEX10 through its zinc-binding RING domain; loss of PEX10 or PEX12 does not reduce PEX5 docking to the peroxisome membrane, placing PEX10 and PEX12 downstream of the receptor-docking step in the peroxisomal matrix protein import pathway.","method":"Two-hybrid assays, blot overlay assays, coimmunoprecipitation, genetic suppression by overexpression, missense patient mutation analysis","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1–2 — multiple orthogonal methods (Y2H, overlay, Co-IP) plus in vivo genetic suppression; highly cited foundational study","pmids":["10562279"],"is_preprint":false},{"year":2000,"finding":"The C-terminal zinc-binding RING domain of PEX10 is critical for its function; missense mutation H290Q in this domain reduces activity and correlates with milder clinical phenotype, while nonsense/frameshift mutations removing only the C-terminal third also attenuate activity, demonstrating the zinc-binding domain is the key functional element.","method":"Functional complementation assay in patient fibroblasts, site-directed mutagenesis of PEX10 cDNA constructs","journal":"Human mutation","confidence":"Medium","confidence_rationale":"Tier 2 — in-cell functional complementation with defined mutations, single lab","pmids":["10862081"],"is_preprint":false},{"year":2025,"finding":"PEX10 forms part of a peroxisomal Pex2/Pex10/Pex12 E3-ubiquitin ligase complex that, upon Pex5-Pex8 complex formation, initiates recycling (ubiquitination) of the PTS1 receptor Pex5 after cargo translocation into peroxisomes.","method":"Structural (cryo-EM/X-ray of Pex8-Pex5 complex), biochemical interaction assays, functional translocation assays in yeast","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 1 — structural and functional data, but preprint not yet peer-reviewed","pmids":["bio_10.1101_2025.08.30.673231"],"is_preprint":true},{"year":2024,"finding":"In prostate cancer cells, PEX10 expression is transcriptionally induced by reactive oxygen species (ROS) activators; the androgen receptor (AR) promotes PEX10 expression in cooperation with FOXA1 as an enhancer. PEX10 reduces intracellular ROS levels, and enzalutamide inhibits PEX10 expression by blocking AR function, thereby sensitizing cells to ROS activators.","method":"ROS assays, AR ChIP/enhancer analysis with FOXA1, enzalutamide treatment with PEX10 knockdown/overexpression in prostate cancer cell lines","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 — multiple cellular assays with mechanistic pathway placement (AR→FOXA1→PEX10→ROS), single lab","pmids":["39097593"],"is_preprint":false},{"year":2006,"finding":"Arabidopsis PEX19 (a cytosolic peroxisomal membrane protein chaperone) physically binds PEX10 through the C-terminus of PEX10, as demonstrated by co-immunoprecipitation and GST pull-down assays, indicating PEX10 is a cargo/client of PEX19 for targeting to the peroxisome membrane.","method":"In vitro translation and co-immunoprecipitation, GST pull-down with recombinant proteins","journal":"Molecular membrane biology","confidence":"Medium","confidence_rationale":"Tier 3 — interaction shown by two binding assays in plant orthologs, mechanistically consistent with mammalian peroxisome biology","pmids":["16923726"],"is_preprint":false}],"current_model":"PEX10 is an integral peroxisomal membrane protein with a cytosol-facing C-terminal C3HC4 zinc RING domain; it acts downstream of PEX5 receptor docking as part of a Pex2/Pex10/Pex12 E3-ubiquitin ligase complex that ubiquitinates the PTS1 receptor PEX5 to drive its recycling after cargo translocation, and loss of PEX10 selectively blocks peroxisomal matrix protein import while leaving membrane protein targeting and peroxisome formation intact."},"narrative":{"teleology":[{"year":1998,"claim":"Identification of PEX10 as the gene mutated in a peroxisome biogenesis disorder complementation group established that it encodes an integral peroxisomal membrane protein whose loss selectively abolishes matrix protein import without affecting membrane biogenesis.","evidence":"Expression of epitope-tagged PEX10 in patient fibroblasts with complementation assays and topology mapping in two independent labs","pmids":["9700193","9683594"],"confidence":"High","gaps":["Position of PEX10 within the import pathway relative to receptor docking was unknown","Functional role of the RING domain had not been tested by mutagenesis","Direct binding partners on the peroxisome membrane were uncharacterized"]},{"year":1999,"claim":"Demonstration that PEX12 physically interacts with both PEX5 and PEX10 through its RING domain, and that PEX10/PEX12 loss does not reduce PEX5 docking, placed PEX10 at a post-docking step in the import cycle.","evidence":"Two-hybrid, blot overlay, co-immunoprecipitation, and genetic suppression assays in mammalian and yeast systems","pmids":["10562279"],"confidence":"High","gaps":["Whether PEX10 directly ubiquitinates PEX5 or acts indirectly was not resolved","Stoichiometry and architecture of the PEX2/PEX10/PEX12 complex were unknown"]},{"year":2000,"claim":"Mutagenesis of the RING zinc-binding domain proved it is the key functional element of PEX10, with a point mutation (H290Q) causing partial loss of activity that correlates with a milder clinical phenotype.","evidence":"Site-directed mutagenesis and functional complementation in patient fibroblasts","pmids":["10862081"],"confidence":"Medium","gaps":["Biochemical activity of the RING domain (E3 ligase activity) was not directly assayed","Single-lab study without independent replication of genotype–phenotype correlation"]},{"year":2006,"claim":"Physical interaction between plant PEX19 and the C-terminus of PEX10 identified PEX19 as the likely chaperone/receptor for PEX10 targeting to the peroxisomal membrane.","evidence":"Co-immunoprecipitation and GST pull-down with recombinant Arabidopsis proteins","pmids":["16923726"],"confidence":"Medium","gaps":["Demonstrated in plant orthologs; direct binding in mammalian system not shown in this study","Whether PEX19 binding is required for PEX10 membrane insertion was not tested"]},{"year":2024,"claim":"Discovery that androgen receptor and FOXA1 cooperatively drive PEX10 transcription in prostate cancer cells, and that PEX10 reduces intracellular ROS, linked peroxisomal biogenesis to cancer-cell redox homeostasis.","evidence":"ROS assays, AR ChIP/enhancer analysis, enzalutamide treatment with PEX10 knockdown/overexpression in prostate cancer cell lines","pmids":["39097593"],"confidence":"Medium","gaps":["Single-lab study; whether AR-driven PEX10 expression is relevant beyond prostate cancer is untested","Whether ROS reduction is a direct consequence of restored peroxisomal import or an indirect effect is unclear"]},{"year":null,"claim":"The precise catalytic contribution of PEX10 within the Pex2/Pex10/Pex12 RING complex—whether PEX10 directly transfers ubiquitin to PEX5 or serves a structural/scaffolding role—remains unresolved, as does a high-resolution structure of the mammalian complex.","evidence":"","pmids":[],"confidence":"High","gaps":["No in vitro ubiquitin transfer assay with purified mammalian PEX10 has been reported","High-resolution structure of the human Pex2/Pex10/Pex12 complex is lacking","Regulation of complex assembly and turnover during the import cycle is not understood"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,2,3]}],"localization":[{"term_id":"GO:0043226","term_label":"organelle","supporting_discovery_ids":[0,1,5]}],"pathway":[{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[0,1,3]},{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[0,1]}],"complexes":["Pex2/Pex10/Pex12 E3 ligase complex"],"partners":["PEX12","PEX5","PEX19","PEX2"],"other_free_text":[]},"mechanistic_narrative":"PEX10 is an integral peroxisomal membrane protein with two transmembrane segments and a cytosol-facing C3HC4 zinc RING domain that functions as a core component of the Pex2/Pex10/Pex12 E3-ubiquitin ligase complex required for peroxisomal matrix protein import [PMID:9700193, PMID:9683594]. PEX10 acts downstream of PTS1 receptor (PEX5) docking at the peroxisomal membrane; loss of PEX10 does not impair PEX5 membrane association but blocks matrix protein translocation, while peroxisome membrane biogenesis remains intact [PMID:10562279]. The RING domain is the essential functional element: missense mutations within it attenuate complementation activity, and the domain mediates physical interaction with PEX12 to coordinate PEX5 ubiquitination and recycling after cargo delivery [PMID:10862081, PMID:10562279]. Biallelic loss-of-function mutations in PEX10 cause peroxisome biogenesis disorders of the Zellweger spectrum [PMID:9700193]."},"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":115,"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":88,"is_preprint":false},{"pmid":"14576288","id":"PMC_14576288","title":"An Arabidopsis pex10 null mutant is embryo lethal, implicating peroxisomes in an essential role during plant embryogenesis.","date":"2003","source":"Plant physiology","url":"https://pubmed.ncbi.nlm.nih.gov/14576288","citation_count":88,"is_preprint":false},{"pmid":"17215364","id":"PMC_17215364","title":"Requirement of the C3HC4 zinc RING finger of the Arabidopsis PEX10 for photorespiration and leaf peroxisome contact with chloroplasts.","date":"2007","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/17215364","citation_count":76,"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":47,"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":31,"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":"16923726","id":"PMC_16923726","title":"Arabidopsis PEX19 is a dimeric protein that binds the peroxin PEX10.","date":"2006","source":"Molecular membrane biology","url":"https://pubmed.ncbi.nlm.nih.gov/16923726","citation_count":21,"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":1,"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":13248,"output_tokens":1560,"usd":0.031572},"stage2":{"model":"claude-opus-4-6","input_tokens":4810,"output_tokens":1749,"usd":0.101663},"total_usd":0.133235,"stage1_batch_id":"msgbatch_012ZiiCUBfB62NCTQd4Bk1Jo","stage2_batch_id":"msgbatch_01Xuqq9aUHoQouS7UVGZNStZ","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. Loss of PEX10 function blocks peroxisomal matrix protein import while peroxisome membrane protein import and peroxisome formation remain intact.\",\n      \"method\": \"Expression of epitope-tagged PEX10 in patient fibroblasts, functional complementation assay, mutation analysis\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — topology mapping by expression of epitope-tagged protein, functional complementation with defined mutations, replicated across two independent labs (PMID:9700193 and PMID:9683594)\",\n      \"pmids\": [\"9700193\", \"9683594\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"PEX12 physically interacts with both PEX5 (PTS1 receptor) and PEX10 through its zinc-binding RING domain; loss of PEX10 or PEX12 does not reduce PEX5 docking to the peroxisome membrane, placing PEX10 and PEX12 downstream of the receptor-docking step in the peroxisomal matrix protein import pathway.\",\n      \"method\": \"Two-hybrid assays, blot overlay assays, coimmunoprecipitation, genetic suppression by overexpression, missense patient mutation analysis\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods (Y2H, overlay, Co-IP) plus in vivo genetic suppression; highly cited foundational study\",\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 its function; missense mutation H290Q in this domain reduces activity and correlates with milder clinical phenotype, while nonsense/frameshift mutations removing only the C-terminal third also attenuate activity, demonstrating the zinc-binding domain is the key functional element.\",\n      \"method\": \"Functional complementation assay in patient fibroblasts, site-directed mutagenesis of PEX10 cDNA constructs\",\n      \"journal\": \"Human mutation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in-cell functional complementation with defined mutations, single lab\",\n      \"pmids\": [\"10862081\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PEX10 forms part of a peroxisomal Pex2/Pex10/Pex12 E3-ubiquitin ligase complex that, upon Pex5-Pex8 complex formation, initiates recycling (ubiquitination) of the PTS1 receptor Pex5 after cargo translocation into peroxisomes.\",\n      \"method\": \"Structural (cryo-EM/X-ray of Pex8-Pex5 complex), biochemical interaction assays, functional translocation assays in yeast\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 — structural and functional data, but preprint not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.08.30.673231\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In prostate cancer cells, PEX10 expression is transcriptionally induced by reactive oxygen species (ROS) activators; the androgen receptor (AR) promotes PEX10 expression in cooperation with FOXA1 as an enhancer. PEX10 reduces intracellular ROS levels, and enzalutamide inhibits PEX10 expression by blocking AR function, thereby sensitizing cells to ROS activators.\",\n      \"method\": \"ROS assays, AR ChIP/enhancer analysis with FOXA1, enzalutamide treatment with PEX10 knockdown/overexpression in prostate cancer cell lines\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple cellular assays with mechanistic pathway placement (AR→FOXA1→PEX10→ROS), single lab\",\n      \"pmids\": [\"39097593\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Arabidopsis PEX19 (a cytosolic peroxisomal membrane protein chaperone) physically binds PEX10 through the C-terminus of PEX10, as demonstrated by co-immunoprecipitation and GST pull-down assays, indicating PEX10 is a cargo/client of PEX19 for targeting to the peroxisome membrane.\",\n      \"method\": \"In vitro translation and co-immunoprecipitation, GST pull-down with recombinant proteins\",\n      \"journal\": \"Molecular membrane biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — interaction shown by two binding assays in plant orthologs, mechanistically consistent with mammalian peroxisome biology\",\n      \"pmids\": [\"16923726\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"PEX10 is an integral peroxisomal membrane protein with a cytosol-facing C-terminal C3HC4 zinc RING domain; it acts downstream of PEX5 receptor docking as part of a Pex2/Pex10/Pex12 E3-ubiquitin ligase complex that ubiquitinates the PTS1 receptor PEX5 to drive its recycling after cargo translocation, and loss of PEX10 selectively blocks peroxisomal matrix protein import while leaving membrane protein targeting and peroxisome formation intact.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"PEX10 is an integral peroxisomal membrane protein with two transmembrane segments and a cytosol-facing C3HC4 zinc RING domain that functions as a core component of the Pex2/Pex10/Pex12 E3-ubiquitin ligase complex required for peroxisomal matrix protein import [PMID:9700193, PMID:9683594]. PEX10 acts downstream of PTS1 receptor (PEX5) docking at the peroxisomal membrane; loss of PEX10 does not impair PEX5 membrane association but blocks matrix protein translocation, while peroxisome membrane biogenesis remains intact [PMID:10562279]. The RING domain is the essential functional element: missense mutations within it attenuate complementation activity, and the domain mediates physical interaction with PEX12 to coordinate PEX5 ubiquitination and recycling after cargo delivery [PMID:10862081, PMID:10562279]. Biallelic loss-of-function mutations in PEX10 cause peroxisome biogenesis disorders of the Zellweger spectrum [PMID:9700193].\",\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Identification of PEX10 as the gene mutated in a peroxisome biogenesis disorder complementation group established that it encodes an integral peroxisomal membrane protein whose loss selectively abolishes matrix protein import without affecting membrane biogenesis.\",\n      \"evidence\": \"Expression of epitope-tagged PEX10 in patient fibroblasts with complementation assays and topology mapping in two independent labs\",\n      \"pmids\": [\"9700193\", \"9683594\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Position of PEX10 within the import pathway relative to receptor docking was unknown\",\n        \"Functional role of the RING domain had not been tested by mutagenesis\",\n        \"Direct binding partners on the peroxisome membrane were uncharacterized\"\n      ]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Demonstration that PEX12 physically interacts with both PEX5 and PEX10 through its RING domain, and that PEX10/PEX12 loss does not reduce PEX5 docking, placed PEX10 at a post-docking step in the import cycle.\",\n      \"evidence\": \"Two-hybrid, blot overlay, co-immunoprecipitation, and genetic suppression assays in mammalian and yeast systems\",\n      \"pmids\": [\"10562279\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether PEX10 directly ubiquitinates PEX5 or acts indirectly was not resolved\",\n        \"Stoichiometry and architecture of the PEX2/PEX10/PEX12 complex were unknown\"\n      ]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Mutagenesis of the RING zinc-binding domain proved it is the key functional element of PEX10, with a point mutation (H290Q) causing partial loss of activity that correlates with a milder clinical phenotype.\",\n      \"evidence\": \"Site-directed mutagenesis and functional complementation in patient fibroblasts\",\n      \"pmids\": [\"10862081\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Biochemical activity of the RING domain (E3 ligase activity) was not directly assayed\",\n        \"Single-lab study without independent replication of genotype–phenotype correlation\"\n      ]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Physical interaction between plant PEX19 and the C-terminus of PEX10 identified PEX19 as the likely chaperone/receptor for PEX10 targeting to the peroxisomal membrane.\",\n      \"evidence\": \"Co-immunoprecipitation and GST pull-down with recombinant Arabidopsis proteins\",\n      \"pmids\": [\"16923726\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Demonstrated in plant orthologs; direct binding in mammalian system not shown in this study\",\n        \"Whether PEX19 binding is required for PEX10 membrane insertion was not tested\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Discovery that androgen receptor and FOXA1 cooperatively drive PEX10 transcription in prostate cancer cells, and that PEX10 reduces intracellular ROS, linked peroxisomal biogenesis to cancer-cell redox homeostasis.\",\n      \"evidence\": \"ROS assays, AR ChIP/enhancer analysis, enzalutamide treatment with PEX10 knockdown/overexpression in prostate cancer cell lines\",\n      \"pmids\": [\"39097593\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Single-lab study; whether AR-driven PEX10 expression is relevant beyond prostate cancer is untested\",\n        \"Whether ROS reduction is a direct consequence of restored peroxisomal import or an indirect effect is unclear\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The precise catalytic contribution of PEX10 within the Pex2/Pex10/Pex12 RING complex—whether PEX10 directly transfers ubiquitin to PEX5 or serves a structural/scaffolding role—remains unresolved, as does a high-resolution structure of the mammalian complex.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No in vitro ubiquitin transfer assay with purified mammalian PEX10 has been reported\",\n        \"High-resolution structure of the human Pex2/Pex10/Pex12 complex is lacking\",\n        \"Regulation of complex assembly and turnover during the import cycle is not understood\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 2, 3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0043226\", \"supporting_discovery_ids\": [0, 1, 5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [0, 1, 3]},\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"complexes\": [\n      \"Pex2/Pex10/Pex12 E3 ligase complex\"\n    ],\n    \"partners\": [\n      \"PEX12\",\n      \"PEX5\",\n      \"PEX19\",\n      \"PEX2\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}