{"gene":"RHBDF1","run_date":"2026-04-28T19:45:45","timeline":{"discoveries":[{"year":2005,"finding":"RHBDF1 (p100hRho) is a seven-transmembrane protein with a long N-terminal cytoplasmic extension, localizes to the endoplasmic reticulum and Golgi (not cell surface), forms dimers, and interacts with TGF-alpha ligands through a luminal interaction with the EGF core ectodomain. It lacks critical residues for serine protease activity and is thus catalytically inactive.","method":"cDNA cloning, subcellular fractionation/localization, co-immunoprecipitation, active-site sequence analysis, Drosophila functional assay","journal":"Developmental Dynamics","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (localization, co-IP, sequence analysis, in vivo functional assay) in a single study","pmids":["15965977"],"is_preprint":false},{"year":2008,"finding":"RHBDF1 localizes mainly in the endoplasmic reticulum and participates in GPCR-mediated EGFR transactivation by promoting GRP-stimulated secretion (shedding) of TGF-alpha, without affecting production of latent TGF-alpha; RHBDF1 silencing blocks GRP-induced phosphorylation of EGFR, p44/42 MAPK, and AKT while leaving direct EGF-stimulated EGFR signaling intact.","method":"siRNA knockdown, overexpression, EGFR/MAPK/AKT phosphorylation assays, TGF-alpha secretion assay, subcellular localization","journal":"FASEB Journal","confidence":"High","confidence_rationale":"Tier 2 — reciprocal gain/loss-of-function with defined molecular readouts and pathway placement","pmids":["18832597"],"is_preprint":false},{"year":2015,"finding":"iRhom1 (RHBDF1) mediates intracellular transport and maturation of ADAM17; deletion of the extended amino-terminal cytoplasmic domain of iRhom1 increases ADAM17 activity and TNFR shedding, demonstrating that the N-terminal cytoplasmic domain negatively regulates ADAM17 activity. iRhom1 and iRhom2 are functionally redundant in this context.","method":"Genetic screen with ΔN mutants, ADAM17 inhibitor experiments, TNFR shedding assays, cell death assays","journal":"Science Signaling","confidence":"High","confidence_rationale":"Tier 2 — genetic screen plus pharmacological rescue with defined molecular phenotype","pmids":["26535007"],"is_preprint":false},{"year":2015,"finding":"iRhom1 (RHBDF1) stimulates proteasome activity by interacting with 20S proteasome assembly chaperones PAC1 and PAC2, stabilizing them and promoting their dimerization; iRhom1 expression is upregulated by ER stressors, leading to enhanced proteasome activity particularly in ER-containing microsomes.","method":"Genome-wide cDNA functional screen, siRNA knockdown, overexpression, native-gel and fractionation analysis, co-immunoprecipitation, Drosophila in vivo rescue assay","journal":"Scientific Reports","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods including co-IP, enzymatic activity assays, and in vivo rescue","pmids":["26109405"],"is_preprint":false},{"year":2020,"finding":"Endogenous iRhom1 (RHBDF1) is present on the cell surface (shown by cell-surface biotinylation). Unlike iRhom2, the stability of iRhom1 does not depend on ADAM17 — iRhom1 levels are slightly increased rather than reduced in ADAM17-deficient mouse embryonic fibroblasts, indicating iRhom1 and ADAM17 are not obligate stabilizing partners.","method":"Cell-surface biotinylation of endogenous proteins, ADAM17-knockout mouse embryonic fibroblasts, western blot","journal":"Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — clean KO with defined molecular phenotype but single lab","pmids":["32060096"],"is_preprint":false},{"year":2020,"finding":"In endothelial cells, iRhom1 (RHBDF1) is upregulated by physiological shear stress (partially via transcription factor KLF2), while iRhom2 is upregulated by inflammatory cytokines via NFκB/AP-1; shear stress-driven iRhom1 upregulation affects ADAM17 maturation and surface expression independently of inflammatory stimulation.","method":"Primary endothelial cell culture with shear stress and cytokine stimulation, qPCR, transcriptional inhibitors, ADAM17 maturation and JAM-A shedding assays","journal":"Frontiers in Cardiovascular Medicine","confidence":"Medium","confidence_rationale":"Tier 2 — mechanistic pathway identified with multiple conditions but single lab","pmids":["33335915"],"is_preprint":false},{"year":2021,"finding":"iRhom1 (RHBDF1) controls ectodomain shedding of membrane proteins in the nervous system through ADAM17; proteomic secretome analysis of iRhom1-deficient mouse neurons and cerebrospinal fluid identified MEGF10 as an iRhom1-dependent ADAM17 substrate, validated further using ADAM17-deficient mouse embryonic fibroblasts.","method":"iRhom1-knockout mouse model, hiSPECS secretome proteomics, CSF proteomics, ADAM17-KO MEF validation","journal":"FASEB Journal","confidence":"High","confidence_rationale":"Tier 2 — in vivo KO model with proteomics and orthogonal genetic validation","pmids":["34613632"],"is_preprint":false},{"year":2022,"finding":"An alternative splicing variant of RHBDF1 (RHX6) antagonizes RHBDF1 activity by inhibiting ADAM17/TACE maturation and blocking intracellular transport of pro-TGF-alpha to the cell surface, thereby inhibiting EGFR activation; RHX6 production is regulated by the alternative splicing regulator RBM4.","method":"Overexpression of splicing variant, TACE maturation assay, pro-TGF-alpha trafficking assay, proliferation/migration assays, RBM4 knockdown","journal":"Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — multiple functional assays with defined molecular mechanism, single lab","pmids":["35595096"],"is_preprint":false},{"year":2023,"finding":"RHBDF1 directly interacts with BiP (GRP78) and stabilizes BiP protein; RHBDF1 deletion causes aggregation of unfolded proteins near the ER, and the RHBDF1-BiP interaction maintains ER protein homeostasis in breast cancer cells.","method":"Co-immunoprecipitation, RHBDF1 deletion/silencing, unfolded protein aggregation assay, SPR binding assay with derived peptide","journal":"Acta Pharmacologica Sinica","confidence":"Medium","confidence_rationale":"Tier 2 — direct binding shown by co-IP and SPR, functional consequence shown by KO, single lab","pmids":["37798352"],"is_preprint":false},{"year":2023,"finding":"RHBDF1 protects glucose-6-phosphate isomerase (GPI) from TRIM32-mediated K27/K63-linked ubiquitination and lysosomal degradation by competing with GPI for binding to the NHL domain of the E3 ubiquitin ligase TRIM32 via its multi-transmembrane domain; mouse RHBDF1 residues R747 and Y799 are critical for this competitive binding.","method":"Co-immunoprecipitation, ubiquitination assays, mutagenesis of RHBDF1 (R747, Y799), lysosomal degradation assays, in vivo mouse melanoma model","journal":"Pharmacological Research","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP, mutagenesis, and in vivo validation but single lab","pmids":["37979663"],"is_preprint":false},{"year":2024,"finding":"RHBDF1 promotes PERK expression and the PERK/peIF2α UPR pathway via the JNK/FoxO3 axis: RHBDF1 activates JNK, causing FoxO3 to translocate into the nucleus and drive PERK transcription; RHBDF1 deficiency reduces PERK, pPERK, and peIF2α levels.","method":"RHBDF1 knockdown/overexpression, nuclear fractionation of FoxO3, western blotting for PERK/pPERK/peIF2α, JNK activation assay","journal":"Acta Biochimica et Biophysica Sinica","confidence":"Low","confidence_rationale":"Tier 3 — pathway placement without direct mechanistic reconstitution, single lab","pmids":["39420837"],"is_preprint":false},{"year":2024,"finding":"RHBDF1 interacts with YAP1, and this interaction increases Smad2 phosphorylation and promotes Smad2 nuclear translocation, modulating cisplatin sensitivity in small cell lung cancer cells.","method":"Co-immunoprecipitation, gain- and loss-of-function experiments, western blotting for pSmad2, nuclear fractionation","journal":"Heliyon","confidence":"Low","confidence_rationale":"Tier 3 — single co-IP with pathway readout, single lab","pmids":["39027514"],"is_preprint":false},{"year":2024,"finding":"RHBDF1 facilitates nuclear translocation of PKCζ by interacting with both importin β1 and PKCζ and promoting PKCζ phosphorylation, leading to disruption of the Par apicobasal polarity complex, loss of tight/adherens junction proteins, and increased cell mobility in mammary epithelial cells.","method":"Co-immunoprecipitation (RHBDF1 with importin β1 and PKCζ), PKCζ phosphorylation assay, nuclear fractionation, PKCζ inhibitor rescue experiment, overexpression in mammary epithelial cells","journal":"Biological Research","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP, phosphorylation assay, and pharmacological rescue with defined polarity phenotype, single lab","pmids":["39582014"],"is_preprint":false},{"year":2026,"finding":"Cryo-EM structure of the ADAM17 zymogen bound to iRhom1 reveals that a transmembrane α-helix and a conserved cytoplasmic 're-entry loop' in iRhom1 function as a molecular relay transmitting intracellular signals across the membrane to activate ADAM17; a cardiomyopathy-associated human iRhom1 mutation disrupts ADAM17 maturation and trafficking.","method":"Cryo-electron microscopy structure determination, all-atom molecular dynamics simulations, disease-associated mutant functional analysis (ADAM17 maturation/trafficking assay)","journal":"Cell Reports","confidence":"High","confidence_rationale":"Tier 1 — cryo-EM structure with functional validation of key structural elements and disease mutant","pmids":["42024498"],"is_preprint":false}],"current_model":"RHBDF1 (iRhom1) is a catalytically inactive, multi-transmembrane ER-resident pseudoprotease that acts as an essential regulatory subunit of ADAM17: its transmembrane helix and cytoplasmic re-entry loop form a molecular relay (visualized by cryo-EM) that promotes ADAM17 maturation, trafficking, and activation, thereby controlling shedding of EGFR ligands (e.g., TGF-alpha) and other substrates (TNF receptors, MEGF10); its extended N-terminal cytoplasmic domain negatively regulates ADAM17 activity, its stability is ADAM17-independent (unlike iRhom2), and it additionally engages BiP to maintain ER proteostasis, interacts with PAC1/PAC2 to stimulate proteasome assembly under ER stress, competes with GPI for TRIM32-mediated ubiquitination to regulate glycolysis, and facilitates PKCζ nuclear translocation via importin β1 to disrupt epithelial apicobasal polarity."},"narrative":{"teleology":[{"year":2005,"claim":"Identification of RHBDF1 as a catalytically dead, ER/Golgi-resident rhomboid-family protein that dimerizes and physically engages TGF-α established the gene as a pseudoprotease with a potential role in EGFR ligand processing.","evidence":"cDNA cloning, subcellular fractionation, co-immunoprecipitation, active-site sequence analysis, and Drosophila functional assay","pmids":["15965977"],"confidence":"High","gaps":["Whether RHBDF1 promotes TGF-α shedding or only intracellular trafficking was unresolved","Identity of the metalloprotease partner was unknown","No structure of the RHBDF1 transmembrane domain"]},{"year":2008,"claim":"Demonstrating that RHBDF1 is required for GPCR-induced TGF-α shedding and subsequent EGFR transactivation positioned it as a specific gatekeeper of ligand release rather than ligand production.","evidence":"siRNA knockdown and overexpression in cells with EGFR/MAPK/AKT phosphorylation readouts and TGF-α secretion assays","pmids":["18832597"],"confidence":"High","gaps":["The shedding enzyme mediating TGF-α release downstream of RHBDF1 was not identified","Whether RHBDF1 acts at the ER or at the cell surface was unclear"]},{"year":2015,"claim":"Linking RHBDF1 directly to ADAM17 maturation and showing that the N-terminal cytoplasmic domain is an inhibitory module unified prior observations into a model where iRhom1 is an essential ADAM17 regulatory subunit, while the parallel discovery of PAC1/PAC2 interaction revealed an ADAM17-independent role in ER-stress-induced proteasome assembly.","evidence":"ΔN-mutant genetic screen with ADAM17 inhibitor rescue and TNFR shedding assays; genome-wide cDNA screen with co-IP, native-gel proteasome analysis, and Drosophila rescue","pmids":["26535007","26109405"],"confidence":"High","gaps":["Structural basis of the iRhom1–ADAM17 interaction was unknown","Whether proteasome-stimulatory and ADAM17-regulatory functions are coordinated was untested","Redundancy with iRhom2 complicated tissue-specific loss-of-function interpretation"]},{"year":2020,"claim":"Showing that endogenous iRhom1 reaches the cell surface and that its stability is ADAM17-independent (unlike iRhom2) clarified that iRhom1 has autonomous functions and distinct regulation from its paralog; transcriptional induction by shear stress via KLF2 placed it in vascular mechano-sensing.","evidence":"Cell-surface biotinylation in ADAM17-KO MEFs; primary endothelial cells under shear stress with qPCR and ADAM17 maturation assays","pmids":["32060096","33335915"],"confidence":"Medium","gaps":["Both findings from single laboratories, awaiting independent replication","Function of surface-localized iRhom1 pool was not determined","Downstream vascular phenotype of iRhom1 loss in shear-stress context was unexplored"]},{"year":2021,"claim":"In vivo secretome profiling of iRhom1-knockout mouse neurons identified MEGF10 as a physiological, nervous-system-specific ADAM17 substrate controlled by iRhom1, extending its functional scope beyond EGFR ligands.","evidence":"iRhom1-KO mouse model with hiSPECS secretome proteomics, CSF proteomics, and ADAM17-KO MEF validation","pmids":["34613632"],"confidence":"High","gaps":["Functional consequence of reduced MEGF10 shedding in the CNS was not addressed","Full catalogue of iRhom1-specific versus iRhom2-dependent substrates in vivo remains incomplete"]},{"year":2022,"claim":"Discovery of the alternative splice variant RHX6 that antagonizes RHBDF1 by blocking ADAM17 maturation and TGF-α trafficking revealed a built-in post-transcriptional brake on iRhom1 function, regulated by the splicing factor RBM4.","evidence":"Overexpression of RHX6 splicing variant with TACE maturation, pro-TGF-α trafficking, and proliferation/migration assays; RBM4 knockdown","pmids":["35595096"],"confidence":"Medium","gaps":["Physiological tissue contexts where RHX6 predominates are not defined","Mechanism by which RHX6 interferes with ADAM17 at the molecular level is not resolved"]},{"year":2023,"claim":"Two ADAM17-independent functions were delineated: RHBDF1 stabilizes BiP to maintain ER protein homeostasis, and it competitively shields GPI from TRIM32-mediated K27/K63 ubiquitination to sustain glycolysis, broadening its role to metabolic and proteostasis regulation.","evidence":"Co-IP and SPR for BiP binding with RHBDF1-KO unfolded protein aggregation assay; co-IP, ubiquitination assays, mutagenesis (R747, Y799), and in vivo mouse melanoma model for TRIM32/GPI axis","pmids":["37798352","37979663"],"confidence":"Medium","gaps":["BiP interaction and TRIM32 competition each demonstrated in single labs","Whether these functions depend on iRhom1 pseudoprotease fold or only its transmembrane domain is unknown","Relative importance of ADAM17-dependent versus -independent functions in normal tissue physiology is unclear"]},{"year":2024,"claim":"Identification of RHBDF1 as a facilitator of PKCζ nuclear translocation via importin β1, leading to disruption of apicobasal polarity, provided a mechanistic route linking RHBDF1 to epithelial-to-mesenchymal transition-like phenotypes.","evidence":"Co-IP of RHBDF1 with importin β1 and PKCζ, nuclear fractionation, PKCζ inhibitor rescue, and polarity/junction marker analysis in mammary epithelial cells","pmids":["39582014"],"confidence":"Medium","gaps":["Single-lab study; independent confirmation needed","Whether PKCζ transport function is separable from ADAM17 regulatory function is unknown","In vivo relevance for mammary tissue polarity not demonstrated"]},{"year":2026,"claim":"The cryo-EM structure of the ADAM17 zymogen–iRhom1 complex revealed the transmembrane helix and cytoplasmic re-entry loop as a signal relay, and a cardiomyopathy-associated human iRhom1 mutation that disrupts ADAM17 maturation linked the structural mechanism to human disease.","evidence":"Cryo-EM structure determination, all-atom MD simulations, disease-associated mutant analysis of ADAM17 maturation/trafficking","pmids":["42024498"],"confidence":"High","gaps":["Full-length structure including the N-terminal inhibitory domain is lacking","Structural basis for iRhom1-specific versus iRhom2-specific substrate selectivity remains unresolved","Whether the cardiomyopathy phenotype is solely ADAM17-dependent or involves other iRhom1 partners is not established"]},{"year":null,"claim":"How iRhom1 coordinates its multiple partner interactions (ADAM17, BiP, PAC1/PAC2, TRIM32/GPI, importin β1/PKCζ) spatiotemporally within the ER and beyond, and which functions dominate in specific tissues, remains an open question.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No integrative study has examined all iRhom1 functions in a single model system","Full-length iRhom1 structure including the disordered N-terminal domain is needed","Conditional tissue-specific knockout phenotyping beyond the nervous system is limited"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,6,13]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[1,7,12]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[0,1,3,8]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[0]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[4]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,2,7,13]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[3,8,10]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[2,3,9]},{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[2,6,12]}],"complexes":["ADAM17-iRhom1 complex"],"partners":["ADAM17","TGFA","HSPA5","PSMG1","PSMG2","TRIM32","PRKCZ","KPNB1"],"other_free_text":[]},"mechanistic_narrative":"RHBDF1 (iRhom1) is a catalytically inactive rhomboid pseudoprotease that functions as a central regulatory cofactor of ADAM17 and an integrator of ER proteostasis. Its transmembrane helix and a conserved cytoplasmic re-entry loop form a molecular relay—resolved by cryo-EM—that promotes ADAM17 maturation, ER-to-surface trafficking, and sheddase activation, controlling release of EGFR ligands (TGF-α), TNF receptors, and neuronal substrates such as MEGF10, while its extended N-terminal cytoplasmic domain negatively tunes ADAM17 activity [PMID:15965977, PMID:26535007, PMID:34613632, PMID:42024498]. Beyond ADAM17 regulation, RHBDF1 maintains ER homeostasis by stabilizing the chaperone BiP and by promoting proteasome assembly through interaction with PAC1/PAC2, and it modulates glycolysis by competitively shielding GPI from TRIM32-mediated ubiquitination [PMID:26109405, PMID:37798352, PMID:37979663]. RHBDF1 additionally facilitates PKCζ nuclear import via importin β1, disrupting the Par apicobasal polarity complex and tight/adherens junctions in epithelial cells [PMID:39582014]."},"prefetch_data":{"uniprot":{"accession":"Q96CC6","full_name":"Inactive rhomboid protein 1","aliases":["Epidermal growth factor receptor-related protein","Rhomboid 5 homolog 1","Rhomboid family member 1","p100hRho"],"length_aa":855,"mass_kda":97.4,"function":"Regulates ADAM17 protease, a sheddase of the epidermal growth factor (EGF) receptor ligands and TNF, thereby plays a role in sleep, cell survival, proliferation, migration and inflammation. Does not exhibit any protease activity on its own","subcellular_location":"Endoplasmic reticulum membrane; Golgi apparatus membrane","url":"https://www.uniprot.org/uniprotkb/Q96CC6/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RHBDF1","classification":"Not Classified","n_dependent_lines":152,"n_total_lines":1208,"dependency_fraction":0.12582781456953643},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RHBDF1","total_profiled":1310},"omim":[{"mim_id":"618337","title":"FERM DOMAIN-CONTAINING PROTEIN 8; FRMD8","url":"https://www.omim.org/entry/618337"},{"mim_id":"614404","title":"RHOMBOID 5 HOMOLOG 2; RHBDF2","url":"https://www.omim.org/entry/614404"},{"mim_id":"614403","title":"RHOMBOID 5 HOMOLOG 1; RHBDF1","url":"https://www.omim.org/entry/614403"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Uncertain","locations":[{"location":"Vesicles","reliability":"Uncertain"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/RHBDF1"},"hgnc":{"alias_symbol":["EGFR-RS","FLJ2235","Dist1","iRhom1"],"prev_symbol":["C16orf8"]},"alphafold":{"accession":"Q96CC6","domains":[{"cath_id":"1.20.1540.10","chopping":"404-427_653-834","consensus_level":"high","plddt":91.2642,"start":404,"end":834},{"cath_id":"-","chopping":"480-638","consensus_level":"high","plddt":88.2921,"start":480,"end":638}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96CC6","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q96CC6-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q96CC6-F1-predicted_aligned_error_v6.png","plddt_mean":68.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RHBDF1","jax_strain_url":"https://www.jax.org/strain/search?query=RHBDF1"},"sequence":{"accession":"Q96CC6","fasta_url":"https://rest.uniprot.org/uniprotkb/Q96CC6.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q96CC6/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96CC6"}},"corpus_meta":[{"pmid":"18832597","id":"PMC_18832597","title":"Human rhomboid family-1 gene RHBDF1 participates in GPCR-mediated transactivation of EGFR growth signals in head and neck squamous cancer cells.","date":"2008","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/18832597","citation_count":67,"is_preprint":false},{"pmid":"26535007","id":"PMC_26535007","title":"Deletions in the cytoplasmic domain of iRhom1 and iRhom2 promote shedding of the TNF receptor by the protease ADAM17.","date":"2015","source":"Science signaling","url":"https://pubmed.ncbi.nlm.nih.gov/26535007","citation_count":50,"is_preprint":false},{"pmid":"15965977","id":"PMC_15965977","title":"Characterization of a human rhomboid homolog, p100hRho/RHBDF1, which interacts with TGF-alpha family ligands.","date":"2005","source":"Developmental dynamics : an official publication of the American Association of Anatomists","url":"https://pubmed.ncbi.nlm.nih.gov/15965977","citation_count":37,"is_preprint":false},{"pmid":"38177179","id":"PMC_38177179","title":"Inhibition of iRhom1 by CD44-targeting nanocarrier for improved cancer immunochemotherapy.","date":"2024","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/38177179","citation_count":34,"is_preprint":false},{"pmid":"26109405","id":"PMC_26109405","title":"iRhom1 regulates proteasome activity via PAC1/2 under ER stress.","date":"2015","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26109405","citation_count":30,"is_preprint":false},{"pmid":"29654741","id":"PMC_29654741","title":"RHBDF1 regulates APC-mediated stimulation of the epithelial-to-mesenchymal transition and proliferation of colorectal cancer cells in part via the Wnt/β-catenin signalling pathway.","date":"2018","source":"Experimental cell 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glucose-6-phosphate isomerase degradation via TRIM32.","date":"2023","source":"Pharmacological research","url":"https://pubmed.ncbi.nlm.nih.gov/37979663","citation_count":6,"is_preprint":false},{"pmid":"35595096","id":"PMC_35595096","title":"Alternative splicing of the human rhomboid family-1 gene RHBDF1 inhibits epidermal growth factor receptor activation.","date":"2022","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/35595096","citation_count":4,"is_preprint":false},{"pmid":"37798352","id":"PMC_37798352","title":"Pentapeptide PYRAE triggers ER stress-mediated apoptosis of breast cancer cells in mice by targeting RHBDF1-BiP interaction.","date":"2023","source":"Acta pharmacologica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/37798352","citation_count":3,"is_preprint":false},{"pmid":"39420837","id":"PMC_39420837","title":"RHBDF1 promotes PERK expression through the JNK/FoxO3 pathway in breast cancer cells.","date":"2024","source":"Acta biochimica et biophysica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/39420837","citation_count":1,"is_preprint":false},{"pmid":"39027514","id":"PMC_39027514","title":"RHBDF1 modulates cisplatin sensitivity of small cell lung cancer through YAP1/Smad2 signaling pathway.","date":"2024","source":"Heliyon","url":"https://pubmed.ncbi.nlm.nih.gov/39027514","citation_count":1,"is_preprint":false},{"pmid":"39582014","id":"PMC_39582014","title":"Perturbation of mammary epithelial cell apicobasal polarity by RHBDF1-facilitated nuclear translocation of PKCζ.","date":"2024","source":"Biological research","url":"https://pubmed.ncbi.nlm.nih.gov/39582014","citation_count":1,"is_preprint":false},{"pmid":"42024498","id":"PMC_42024498","title":"Structural basis for ADAM17 activation by the iRhom1 pseudoprotease.","date":"2026","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/42024498","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.08.16.24312138","title":"Common and rare variant analyses reveal novel genetic factors underlying Idiopathic Pulmonary Fibrosis and shared aetiology with COVID-19","date":"2024-08-17","source":"bioRxiv","url":"https://doi.org/10.1101/2024.08.16.24312138","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11320,"output_tokens":3566,"usd":0.043725},"stage2":{"model":"claude-opus-4-6","input_tokens":7009,"output_tokens":3098,"usd":0.168742},"total_usd":0.212467,"stage1_batch_id":"msgbatch_01SUR8NDzVrNmyszWyJeCct7","stage2_batch_id":"msgbatch_01G7Yfutko3cDWzejrwH5AMx","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2005,\n      \"finding\": \"RHBDF1 (p100hRho) is a seven-transmembrane protein with a long N-terminal cytoplasmic extension, localizes to the endoplasmic reticulum and Golgi (not cell surface), forms dimers, and interacts with TGF-alpha ligands through a luminal interaction with the EGF core ectodomain. It lacks critical residues for serine protease activity and is thus catalytically inactive.\",\n      \"method\": \"cDNA cloning, subcellular fractionation/localization, co-immunoprecipitation, active-site sequence analysis, Drosophila functional assay\",\n      \"journal\": \"Developmental Dynamics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (localization, co-IP, sequence analysis, in vivo functional assay) in a single study\",\n      \"pmids\": [\"15965977\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"RHBDF1 localizes mainly in the endoplasmic reticulum and participates in GPCR-mediated EGFR transactivation by promoting GRP-stimulated secretion (shedding) of TGF-alpha, without affecting production of latent TGF-alpha; RHBDF1 silencing blocks GRP-induced phosphorylation of EGFR, p44/42 MAPK, and AKT while leaving direct EGF-stimulated EGFR signaling intact.\",\n      \"method\": \"siRNA knockdown, overexpression, EGFR/MAPK/AKT phosphorylation assays, TGF-alpha secretion assay, subcellular localization\",\n      \"journal\": \"FASEB Journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal gain/loss-of-function with defined molecular readouts and pathway placement\",\n      \"pmids\": [\"18832597\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"iRhom1 (RHBDF1) mediates intracellular transport and maturation of ADAM17; deletion of the extended amino-terminal cytoplasmic domain of iRhom1 increases ADAM17 activity and TNFR shedding, demonstrating that the N-terminal cytoplasmic domain negatively regulates ADAM17 activity. iRhom1 and iRhom2 are functionally redundant in this context.\",\n      \"method\": \"Genetic screen with ΔN mutants, ADAM17 inhibitor experiments, TNFR shedding assays, cell death assays\",\n      \"journal\": \"Science Signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic screen plus pharmacological rescue with defined molecular phenotype\",\n      \"pmids\": [\"26535007\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"iRhom1 (RHBDF1) stimulates proteasome activity by interacting with 20S proteasome assembly chaperones PAC1 and PAC2, stabilizing them and promoting their dimerization; iRhom1 expression is upregulated by ER stressors, leading to enhanced proteasome activity particularly in ER-containing microsomes.\",\n      \"method\": \"Genome-wide cDNA functional screen, siRNA knockdown, overexpression, native-gel and fractionation analysis, co-immunoprecipitation, Drosophila in vivo rescue assay\",\n      \"journal\": \"Scientific Reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods including co-IP, enzymatic activity assays, and in vivo rescue\",\n      \"pmids\": [\"26109405\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Endogenous iRhom1 (RHBDF1) is present on the cell surface (shown by cell-surface biotinylation). Unlike iRhom2, the stability of iRhom1 does not depend on ADAM17 — iRhom1 levels are slightly increased rather than reduced in ADAM17-deficient mouse embryonic fibroblasts, indicating iRhom1 and ADAM17 are not obligate stabilizing partners.\",\n      \"method\": \"Cell-surface biotinylation of endogenous proteins, ADAM17-knockout mouse embryonic fibroblasts, western blot\",\n      \"journal\": \"Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with defined molecular phenotype but single lab\",\n      \"pmids\": [\"32060096\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"In endothelial cells, iRhom1 (RHBDF1) is upregulated by physiological shear stress (partially via transcription factor KLF2), while iRhom2 is upregulated by inflammatory cytokines via NFκB/AP-1; shear stress-driven iRhom1 upregulation affects ADAM17 maturation and surface expression independently of inflammatory stimulation.\",\n      \"method\": \"Primary endothelial cell culture with shear stress and cytokine stimulation, qPCR, transcriptional inhibitors, ADAM17 maturation and JAM-A shedding assays\",\n      \"journal\": \"Frontiers in Cardiovascular Medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — mechanistic pathway identified with multiple conditions but single lab\",\n      \"pmids\": [\"33335915\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"iRhom1 (RHBDF1) controls ectodomain shedding of membrane proteins in the nervous system through ADAM17; proteomic secretome analysis of iRhom1-deficient mouse neurons and cerebrospinal fluid identified MEGF10 as an iRhom1-dependent ADAM17 substrate, validated further using ADAM17-deficient mouse embryonic fibroblasts.\",\n      \"method\": \"iRhom1-knockout mouse model, hiSPECS secretome proteomics, CSF proteomics, ADAM17-KO MEF validation\",\n      \"journal\": \"FASEB Journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo KO model with proteomics and orthogonal genetic validation\",\n      \"pmids\": [\"34613632\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"An alternative splicing variant of RHBDF1 (RHX6) antagonizes RHBDF1 activity by inhibiting ADAM17/TACE maturation and blocking intracellular transport of pro-TGF-alpha to the cell surface, thereby inhibiting EGFR activation; RHX6 production is regulated by the alternative splicing regulator RBM4.\",\n      \"method\": \"Overexpression of splicing variant, TACE maturation assay, pro-TGF-alpha trafficking assay, proliferation/migration assays, RBM4 knockdown\",\n      \"journal\": \"Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple functional assays with defined molecular mechanism, single lab\",\n      \"pmids\": [\"35595096\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RHBDF1 directly interacts with BiP (GRP78) and stabilizes BiP protein; RHBDF1 deletion causes aggregation of unfolded proteins near the ER, and the RHBDF1-BiP interaction maintains ER protein homeostasis in breast cancer cells.\",\n      \"method\": \"Co-immunoprecipitation, RHBDF1 deletion/silencing, unfolded protein aggregation assay, SPR binding assay with derived peptide\",\n      \"journal\": \"Acta Pharmacologica Sinica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct binding shown by co-IP and SPR, functional consequence shown by KO, single lab\",\n      \"pmids\": [\"37798352\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RHBDF1 protects glucose-6-phosphate isomerase (GPI) from TRIM32-mediated K27/K63-linked ubiquitination and lysosomal degradation by competing with GPI for binding to the NHL domain of the E3 ubiquitin ligase TRIM32 via its multi-transmembrane domain; mouse RHBDF1 residues R747 and Y799 are critical for this competitive binding.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, mutagenesis of RHBDF1 (R747, Y799), lysosomal degradation assays, in vivo mouse melanoma model\",\n      \"journal\": \"Pharmacological Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP, mutagenesis, and in vivo validation but single lab\",\n      \"pmids\": [\"37979663\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RHBDF1 promotes PERK expression and the PERK/peIF2α UPR pathway via the JNK/FoxO3 axis: RHBDF1 activates JNK, causing FoxO3 to translocate into the nucleus and drive PERK transcription; RHBDF1 deficiency reduces PERK, pPERK, and peIF2α levels.\",\n      \"method\": \"RHBDF1 knockdown/overexpression, nuclear fractionation of FoxO3, western blotting for PERK/pPERK/peIF2α, JNK activation assay\",\n      \"journal\": \"Acta Biochimica et Biophysica Sinica\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — pathway placement without direct mechanistic reconstitution, single lab\",\n      \"pmids\": [\"39420837\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RHBDF1 interacts with YAP1, and this interaction increases Smad2 phosphorylation and promotes Smad2 nuclear translocation, modulating cisplatin sensitivity in small cell lung cancer cells.\",\n      \"method\": \"Co-immunoprecipitation, gain- and loss-of-function experiments, western blotting for pSmad2, nuclear fractionation\",\n      \"journal\": \"Heliyon\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single co-IP with pathway readout, single lab\",\n      \"pmids\": [\"39027514\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RHBDF1 facilitates nuclear translocation of PKCζ by interacting with both importin β1 and PKCζ and promoting PKCζ phosphorylation, leading to disruption of the Par apicobasal polarity complex, loss of tight/adherens junction proteins, and increased cell mobility in mammary epithelial cells.\",\n      \"method\": \"Co-immunoprecipitation (RHBDF1 with importin β1 and PKCζ), PKCζ phosphorylation assay, nuclear fractionation, PKCζ inhibitor rescue experiment, overexpression in mammary epithelial cells\",\n      \"journal\": \"Biological Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP, phosphorylation assay, and pharmacological rescue with defined polarity phenotype, single lab\",\n      \"pmids\": [\"39582014\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Cryo-EM structure of the ADAM17 zymogen bound to iRhom1 reveals that a transmembrane α-helix and a conserved cytoplasmic 're-entry loop' in iRhom1 function as a molecular relay transmitting intracellular signals across the membrane to activate ADAM17; a cardiomyopathy-associated human iRhom1 mutation disrupts ADAM17 maturation and trafficking.\",\n      \"method\": \"Cryo-electron microscopy structure determination, all-atom molecular dynamics simulations, disease-associated mutant functional analysis (ADAM17 maturation/trafficking assay)\",\n      \"journal\": \"Cell Reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — cryo-EM structure with functional validation of key structural elements and disease mutant\",\n      \"pmids\": [\"42024498\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RHBDF1 (iRhom1) is a catalytically inactive, multi-transmembrane ER-resident pseudoprotease that acts as an essential regulatory subunit of ADAM17: its transmembrane helix and cytoplasmic re-entry loop form a molecular relay (visualized by cryo-EM) that promotes ADAM17 maturation, trafficking, and activation, thereby controlling shedding of EGFR ligands (e.g., TGF-alpha) and other substrates (TNF receptors, MEGF10); its extended N-terminal cytoplasmic domain negatively regulates ADAM17 activity, its stability is ADAM17-independent (unlike iRhom2), and it additionally engages BiP to maintain ER proteostasis, interacts with PAC1/PAC2 to stimulate proteasome assembly under ER stress, competes with GPI for TRIM32-mediated ubiquitination to regulate glycolysis, and facilitates PKCζ nuclear translocation via importin β1 to disrupt epithelial apicobasal polarity.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"RHBDF1 (iRhom1) is a catalytically inactive rhomboid pseudoprotease that functions as a central regulatory cofactor of ADAM17 and an integrator of ER proteostasis. Its transmembrane helix and a conserved cytoplasmic re-entry loop form a molecular relay—resolved by cryo-EM—that promotes ADAM17 maturation, ER-to-surface trafficking, and sheddase activation, controlling release of EGFR ligands (TGF-α), TNF receptors, and neuronal substrates such as MEGF10, while its extended N-terminal cytoplasmic domain negatively tunes ADAM17 activity [PMID:15965977, PMID:26535007, PMID:34613632, PMID:42024498]. Beyond ADAM17 regulation, RHBDF1 maintains ER homeostasis by stabilizing the chaperone BiP and by promoting proteasome assembly through interaction with PAC1/PAC2, and it modulates glycolysis by competitively shielding GPI from TRIM32-mediated ubiquitination [PMID:26109405, PMID:37798352, PMID:37979663]. RHBDF1 additionally facilitates PKCζ nuclear import via importin β1, disrupting the Par apicobasal polarity complex and tight/adherens junctions in epithelial cells [PMID:39582014].\",\n  \"teleology\": [\n    {\n      \"year\": 2005,\n      \"claim\": \"Identification of RHBDF1 as a catalytically dead, ER/Golgi-resident rhomboid-family protein that dimerizes and physically engages TGF-α established the gene as a pseudoprotease with a potential role in EGFR ligand processing.\",\n      \"evidence\": \"cDNA cloning, subcellular fractionation, co-immunoprecipitation, active-site sequence analysis, and Drosophila functional assay\",\n      \"pmids\": [\"15965977\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether RHBDF1 promotes TGF-α shedding or only intracellular trafficking was unresolved\",\n        \"Identity of the metalloprotease partner was unknown\",\n        \"No structure of the RHBDF1 transmembrane domain\"\n      ]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Demonstrating that RHBDF1 is required for GPCR-induced TGF-α shedding and subsequent EGFR transactivation positioned it as a specific gatekeeper of ligand release rather than ligand production.\",\n      \"evidence\": \"siRNA knockdown and overexpression in cells with EGFR/MAPK/AKT phosphorylation readouts and TGF-α secretion assays\",\n      \"pmids\": [\"18832597\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"The shedding enzyme mediating TGF-α release downstream of RHBDF1 was not identified\",\n        \"Whether RHBDF1 acts at the ER or at the cell surface was unclear\"\n      ]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Linking RHBDF1 directly to ADAM17 maturation and showing that the N-terminal cytoplasmic domain is an inhibitory module unified prior observations into a model where iRhom1 is an essential ADAM17 regulatory subunit, while the parallel discovery of PAC1/PAC2 interaction revealed an ADAM17-independent role in ER-stress-induced proteasome assembly.\",\n      \"evidence\": \"ΔN-mutant genetic screen with ADAM17 inhibitor rescue and TNFR shedding assays; genome-wide cDNA screen with co-IP, native-gel proteasome analysis, and Drosophila rescue\",\n      \"pmids\": [\"26535007\", \"26109405\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis of the iRhom1–ADAM17 interaction was unknown\",\n        \"Whether proteasome-stimulatory and ADAM17-regulatory functions are coordinated was untested\",\n        \"Redundancy with iRhom2 complicated tissue-specific loss-of-function interpretation\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showing that endogenous iRhom1 reaches the cell surface and that its stability is ADAM17-independent (unlike iRhom2) clarified that iRhom1 has autonomous functions and distinct regulation from its paralog; transcriptional induction by shear stress via KLF2 placed it in vascular mechano-sensing.\",\n      \"evidence\": \"Cell-surface biotinylation in ADAM17-KO MEFs; primary endothelial cells under shear stress with qPCR and ADAM17 maturation assays\",\n      \"pmids\": [\"32060096\", \"33335915\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Both findings from single laboratories, awaiting independent replication\",\n        \"Function of surface-localized iRhom1 pool was not determined\",\n        \"Downstream vascular phenotype of iRhom1 loss in shear-stress context was unexplored\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"In vivo secretome profiling of iRhom1-knockout mouse neurons identified MEGF10 as a physiological, nervous-system-specific ADAM17 substrate controlled by iRhom1, extending its functional scope beyond EGFR ligands.\",\n      \"evidence\": \"iRhom1-KO mouse model with hiSPECS secretome proteomics, CSF proteomics, and ADAM17-KO MEF validation\",\n      \"pmids\": [\"34613632\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Functional consequence of reduced MEGF10 shedding in the CNS was not addressed\",\n        \"Full catalogue of iRhom1-specific versus iRhom2-dependent substrates in vivo remains incomplete\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Discovery of the alternative splice variant RHX6 that antagonizes RHBDF1 by blocking ADAM17 maturation and TGF-α trafficking revealed a built-in post-transcriptional brake on iRhom1 function, regulated by the splicing factor RBM4.\",\n      \"evidence\": \"Overexpression of RHX6 splicing variant with TACE maturation, pro-TGF-α trafficking, and proliferation/migration assays; RBM4 knockdown\",\n      \"pmids\": [\"35595096\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Physiological tissue contexts where RHX6 predominates are not defined\",\n        \"Mechanism by which RHX6 interferes with ADAM17 at the molecular level is not resolved\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Two ADAM17-independent functions were delineated: RHBDF1 stabilizes BiP to maintain ER protein homeostasis, and it competitively shields GPI from TRIM32-mediated K27/K63 ubiquitination to sustain glycolysis, broadening its role to metabolic and proteostasis regulation.\",\n      \"evidence\": \"Co-IP and SPR for BiP binding with RHBDF1-KO unfolded protein aggregation assay; co-IP, ubiquitination assays, mutagenesis (R747, Y799), and in vivo mouse melanoma model for TRIM32/GPI axis\",\n      \"pmids\": [\"37798352\", \"37979663\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"BiP interaction and TRIM32 competition each demonstrated in single labs\",\n        \"Whether these functions depend on iRhom1 pseudoprotease fold or only its transmembrane domain is unknown\",\n        \"Relative importance of ADAM17-dependent versus -independent functions in normal tissue physiology is unclear\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identification of RHBDF1 as a facilitator of PKCζ nuclear translocation via importin β1, leading to disruption of apicobasal polarity, provided a mechanistic route linking RHBDF1 to epithelial-to-mesenchymal transition-like phenotypes.\",\n      \"evidence\": \"Co-IP of RHBDF1 with importin β1 and PKCζ, nuclear fractionation, PKCζ inhibitor rescue, and polarity/junction marker analysis in mammary epithelial cells\",\n      \"pmids\": [\"39582014\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Single-lab study; independent confirmation needed\",\n        \"Whether PKCζ transport function is separable from ADAM17 regulatory function is unknown\",\n        \"In vivo relevance for mammary tissue polarity not demonstrated\"\n      ]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"The cryo-EM structure of the ADAM17 zymogen–iRhom1 complex revealed the transmembrane helix and cytoplasmic re-entry loop as a signal relay, and a cardiomyopathy-associated human iRhom1 mutation that disrupts ADAM17 maturation linked the structural mechanism to human disease.\",\n      \"evidence\": \"Cryo-EM structure determination, all-atom MD simulations, disease-associated mutant analysis of ADAM17 maturation/trafficking\",\n      \"pmids\": [\"42024498\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Full-length structure including the N-terminal inhibitory domain is lacking\",\n        \"Structural basis for iRhom1-specific versus iRhom2-specific substrate selectivity remains unresolved\",\n        \"Whether the cardiomyopathy phenotype is solely ADAM17-dependent or involves other iRhom1 partners is not established\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How iRhom1 coordinates its multiple partner interactions (ADAM17, BiP, PAC1/PAC2, TRIM32/GPI, importin β1/PKCζ) spatiotemporally within the ER and beyond, and which functions dominate in specific tissues, remains an open question.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No integrative study has examined all iRhom1 functions in a single model system\",\n        \"Full-length iRhom1 structure including the disordered N-terminal domain is needed\",\n        \"Conditional tissue-specific knockout phenotyping beyond the nervous system is limited\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 6, 13]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [1, 7, 12]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [0, 1, 3, 8]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 2, 7, 13]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [3, 8, 10]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [2, 3, 9]},\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [2, 6, 12]}\n    ],\n    \"complexes\": [\n      \"ADAM17-iRhom1 complex\"\n    ],\n    \"partners\": [\n      \"ADAM17\",\n      \"TGFA\",\n      \"HSPA5\",\n      \"PSMG1\",\n      \"PSMG2\",\n      \"TRIM32\",\n      \"PRKCZ\",\n      \"KPNB1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}