{"gene":"EDEM1","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2003,"finding":"EDEM extracts misfolded glycoproteins (but not productively folding glycoproteins) from the calnexin cycle, promoting their release and accelerating ERAD. EDEM overexpression caused faster release of folding-incompetent proteins from calnexin and earlier onset of degradation; EDEM down-regulation prolonged folding attempts and delayed ERAD.","method":"Overexpression and down-regulation of EDEM combined with pulse-chase analysis of misfolded glycoprotein fate and calnexin association","journal":"Science","confidence":"High","confidence_rationale":"Tier 2 / Strong — two independent labs (PMIDs 12610306 and 12610305) published simultaneously with complementary functional evidence (overexpression, knockdown, co-immunoprecipitation, pulse-chase)","pmids":["12610306","12610305"],"is_preprint":false},{"year":2003,"finding":"EDEM interacts physically with calnexin through its transmembrane region (not with calreticulin). Both binding of substrates to calnexin and their release from calnexin are required for ERAD; EDEM overexpression promoted release of terminally misfolded proteins from calnexin, functioning as an acceptor of calnexin substrates.","method":"Co-immunoprecipitation; overexpression studies with pulse-chase analysis; domain deletion mapping (transmembrane region)","journal":"Science","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP identifying binding partner and domain, replicated finding across two simultaneous independent publications","pmids":["12610305"],"is_preprint":false},{"year":2004,"finding":"EDEM is a soluble protein of the ER lumen (in HEK293 cells), not exclusively an ER membrane protein; it functions as a mannosidase-like chaperone. A second homolog EDEM2 was identified that is also a stress-regulated mannosidase-like protein operating in the ER lumen and accelerates ERAD of terminally misfolded glycoproteins by facilitating their extraction from the calnexin cycle; EDEM2 transcriptional up-regulation depends on the ER stress-activated transcription factor XBP1.","method":"Subcellular fractionation; overexpression and pulse-chase ERAD assays; reporter assays for XBP1-dependent transcription","journal":"The Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — fractionation establishing soluble localization plus functional ERAD assay, single lab","pmids":["15579471"],"is_preprint":false},{"year":2006,"finding":"EDEM1 overexpression accelerates de-mannosylation of terminally misfolded glycoproteins (Man9 or Man5 N-glycans) and inhibits formation of covalent (disulfide-linked) aggregates upon their release from calnexin. Substitution of one conserved catalytic residue in the alpha-1,2-mannosidase active site (E220Q equivalent) fully blocked accelerated de-mannosylation but did not affect the anti-aggregation (chaperone) function of EDEM1, indicating two separable activities.","method":"Pulse-chase with N-glycan analysis; site-directed mutagenesis of conserved catalytic residue; non-reducing SDS-PAGE to detect aggregates","journal":"Biochemical and Biophysical Research Communications","confidence":"High","confidence_rationale":"Tier 1 / Moderate — active-site mutagenesis dissecting two activities (mannosidase vs. chaperone) with multiple orthogonal methods, single lab","pmids":["16987498"],"is_preprint":false},{"year":2006,"finding":"EDEM prevents aberrant covalent dimer formation of misfolded alpha1-antitrypsin NHK, maintaining its retrotranslocation competence. EDEM overexpression selectively prevented accumulation of covalent NHK dimers; this anti-aggregation effect was specific to EDEM and not observed with calnexin or H+/K+-ATPase beta subunit overexpression.","method":"Overexpression of EDEM and control ER membrane proteins; non-reducing SDS-PAGE; pulse-chase degradation assays","journal":"Genes to Cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean functional comparison with controls, single lab","pmids":["16629899"],"is_preprint":false},{"year":2007,"finding":"Endogenous EDEM1 is sequestered in ~150 nm vesicles that bud from rough ER cisternae outside of transitional ER exit sites, lacking a recognizable COPII coat (~87% of immunogold label on vesicles). These vesicles also contain Derlin-2 and misfolded alpha-1-antitrypsin (NHK), indicating a non-canonical ER exit pathway for quality control components and substrates.","method":"High-resolution immunogold electron microscopy and serial section analysis; subcellular fractionation","journal":"Proceedings of the National Academy of Sciences of the USA","confidence":"High","confidence_rationale":"Tier 1 / Moderate — high-resolution structural imaging with quantitative immunogold labeling demonstrating non-COPII vesicle pathway, single lab with rigorous controls","pmids":["17360537"],"is_preprint":false},{"year":2009,"finding":"EDEM1 specifically binds nonnative proteins in a glycan-independent manner. Inhibition of mannosidase activity with kifunensine or mutation of the mannosidase-like domain had no effect on substrate binding but diminished EDEM1 association with the ER membrane adaptor protein SEL1L. EDEM1 thus uses its mannosidase-like domain to target aberrant proteins to the SEL1L-containing dislocation/ubiquitination complex.","method":"Co-immunoprecipitation under glycan-inhibited conditions (kifunensine); mannosidase domain mutants; pulldown assays with SEL1L","journal":"Molecular Cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP, domain mutagenesis, pharmacological inhibition as orthogonal methods, single lab","pmids":["19524542"],"is_preprint":false},{"year":2009,"finding":"EDEM1 functions as a chaperone for rod opsin: it promotes degradation of misfolded P23H rod opsin via ERAD and decreases its aggregation into inclusions. shRNA knockdown of EDEM1 increased P23H rod opsin levels and aggregation. Unexpectedly, EDEM1 binding to rod opsin was independent of mannose trimming, and EDEM1 also promoted cell-surface expression of mutant rod opsin.","method":"shRNA knockdown; overexpression; co-immunoprecipitation; pulse-chase assays; immunofluorescence; endoglycosidase H sensitivity assays","journal":"Journal of Cell Science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function (shRNA) and gain-of-function with multiple readouts, single lab","pmids":["19934218"],"is_preprint":false},{"year":2009,"finding":"Endogenous EDEM1 is degraded by basal autophagy (not by the proteasome under non-starved conditions). EDEM1 was detected in autophagosomes biochemically (LC3 immuno-purification) and by immunocytochemistry. Inhibition of the lysosome-autophagy pathway or knockdown of ATG genes stabilized EDEM1.","method":"Autophagosome immunopurification; immunocytochemistry; pharmacological inhibitors (vinblastine, pepstatin A/E64d, 3-methyladenine); siRNA knockdown of ATGs","journal":"Cellular and Molecular Life Sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal approaches (biochemical fractionation, pharmacology, siRNA), single lab","pmids":["19266160"],"is_preprint":false},{"year":2010,"finding":"EDEM1 overexpression specifically accelerates trimming of alpha-1,2-linked mannose from the C branch of N-glycans, producing Glc1Man8GlcNAc2 isomer C and increasing Man7GlcNAc2 isomer A. The EDEM1 E220Q catalytic mutant did not produce this isomer, confirming alpha-1,2-mannosidase catalytic activity on the C branch. However, ERAD enhancement by wild-type and E220Q EDEM1 was equivalent, attributed to inhibition of aberrant NHK dimer formation.","method":"Radiolabeled N-glycan structural analysis (3H-mannose pulse-chase); site-directed mutagenesis (E220Q); stable cell line overexpression","journal":"Glycobiology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vivo glycan structural analysis with catalytic mutant, rigorous N-glycan mapping, single lab","pmids":["20065073"],"is_preprint":false},{"year":2010,"finding":"Mannose trimming is not required for EDEM1 binding to an ERAD substrate glycoprotein (binding persists with kifunensine treatment or ERManI knockdown). In contrast, substrate association with XTP3-B and E3 ubiquitin ligases HRD1 and SCF(Fbs2) required mannose trimming, placing EDEM1 upstream of these factors in the ERAD pathway.","method":"Co-immunoprecipitation under mannosidase inhibitor (kifunensine) treatment; ERManI siRNA knockdown; colocalization by immunofluorescence at the ERQC","journal":"The Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological and genetic inhibition with co-IP and colocalization, single lab","pmids":["21062743"],"is_preprint":false},{"year":2011,"finding":"EDEM1 overexpression or its up-regulation via IRE1 (unfolded protein response) overrides the mannose-trimming requirement for ERAD, rendering ER mannosidase I dispensable. An EDEM1 deletion mutant lacking most of the carbohydrate-recognition domain still accelerated ERAD and delivered substrate to XTP3-B and OS9. Upon proteasomal inhibition, EDEM1 concentrated with ERAD substrate in the pericentriolar ERQC compartment containing OS9.","method":"ERManI knockdown combined with EDEM1 overexpression; EDEM1 deletion mutants; co-immunoprecipitation; immunofluorescence colocalization at ERQC","journal":"Molecular Biology of the Cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain deletion analysis and epistasis experiments, single lab","pmids":["21917589"],"is_preprint":false},{"year":2011,"finding":"Signal sequence cleavage of EDEM1 is slow and inefficient, producing two isoforms: a soluble form and a type-II membrane form (when signal sequence is uncleaved, creating an N-terminal transmembrane segment). The soluble form efficiently associates with the oxidoreductase ERdj5 and accelerates turnover of soluble ERAD substrates, while the membrane form efficiently associates with SEL1L and accelerates turnover of membrane-associated ERAD substrates.","method":"Signal sequence cleavage analysis; glycosylation site mapping; co-immunoprecipitation with ERdj5 and SEL1L; pulse-chase ERAD assays with isoform-specific constructs","journal":"The Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — biochemical isoform characterization with functional ERAD assays, single lab","pmids":["21632540"],"is_preprint":false},{"year":2012,"finding":"EDEM1 participates in a shared ERAD pathway for both glycosylated and nonglycosylated proteins. Nonglycosylated ERAD substrates (including nonglycosylated H2a, NS-1κ light chain, truncated Igγ heavy chain) co-immunoprecipitate with EDEM1, and require EDEM1 for their degradation. EDEM1 associates with nonglycosylated proteins through a region outside its mannosidase-like domain.","method":"Co-immunoprecipitation; siRNA knockdown of EDEM1; pulse-chase degradation assays; domain deletion analysis","journal":"The Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple substrates, domain mapping, loss-of-function, single lab","pmids":["23233672"],"is_preprint":false},{"year":2012,"finding":"The intrinsically disordered (ID) N-terminal region of EDEM1 (residues ~40–119) mediates binding to both glycosylated and non-glycosylated misfolded proteins. Deletion of this ID region abolished co-immunoprecipitation with misfolded tyrosinase (glycosylated and non-glycosylated mutants), while the intact mannosidase-like domain was not required for substrate binding. EDEM1 overexpression enhanced degradation of wild-type and misfolded tyrosinase.","method":"Co-immunoprecipitation with EDEM1 deletion mutants; homology modeling of mannosidase domain; pulse-chase degradation assays","journal":"PLOS ONE","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain deletion mutants with co-IP and functional assays, single lab","pmids":["22905195"],"is_preprint":false},{"year":2014,"finding":"EDEM1 promotes retrotranslocation of ricin A-chain (RTA) from the ER to the cytosol. EDEM1 overexpression increased RTA retrotranslocation (demonstrated with kifunensine to block competition from misfolded proteins), while RNAi-mediated knockdown decreased RTA retrotranslocation. Co-immunoprecipitation showed that ricin interacts with EDEM1 and with Sec61alpha.","method":"Overexpression and siRNA knockdown of EDEM1; cytosol/ER fractionation to measure retrotranslocation; co-immunoprecipitation","journal":"Molecular Biology of the Cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function and gain-of-function with biochemical fractionation, single lab (replication of PMID 16452630 results)","pmids":["16452630","21388347"],"is_preprint":false},{"year":2014,"finding":"EDEM1 is degraded by selective autophagy. It colocalizes with selective autophagy cargo receptors p62/SQSTM1, NBR1, and Alfy, and is engulfed by autophagic isolation membranes. p62/SQSTM1 and NBR1 knockdown blocked EDEM1 routing to autophagosomes. p62/SQSTM1 interacts only with deglycosylated (and ubiquitinated) EDEM1; deglycosylation by cytosolic peptide N-glycanase (PNGase) is a prerequisite for p62/SQSTM1 interaction and aggregate formation.","method":"Immunofluorescence colocalization; siRNA knockdown of cargo receptors; PNGase inhibitors; ubiquitination assays","journal":"Histochemistry and Cell Biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal approaches (siRNA, inhibitors, imaging), single lab","pmids":["24664425"],"is_preprint":false},{"year":2015,"finding":"Recognition of ERAD substrates by EDEM1 is determined by the hydrophobicity of protein determinants. Mutations increasing or decreasing hydrophobicity in misfolded substrates (ricin A-chain, BACE457) correspondingly altered their interaction with EDEM1. EDEM1 can bind hydrophobic transmembrane regions of misfolded ERAD substrates, and this binding does not require substrate glycosylation.","method":"Site-directed mutagenesis to alter substrate hydrophobicity; co-immunoprecipitation; ERAD assays","journal":"BMC Cell Biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mutagenesis-driven substrate binding analysis, single lab","pmids":["25655076"],"is_preprint":false},{"year":2018,"finding":"EDEM1 and EDEM2 possess intrinsic mannosidase activity in vitro, demonstrated directly. Their activity on free N-glycans and intact glycoproteins is modest, but is significantly enhanced on denatured (unfolded) glycoproteins. EDEM1/2 associate with oxidoreductases including TXNDC11, which enhances their mannosidase activity on glycoproteins but not on free N-glycans.","method":"In vitro mannosidase assay with purified proteins; denatured vs. native substrate comparisons; co-immunoprecipitation with oxidoreductases","journal":"Communications Biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — direct in vitro enzymatic assay establishing catalytic activity and substrate-state dependence, single lab with multiple orthogonal methods","pmids":["30374462"],"is_preprint":false},{"year":2018,"finding":"EDEM1 binds ERAD clients (Z and NHK alpha1-antitrypsin) through a thiol-dependent (redox-sensitive) interaction via Cys256 of the client, as well as through weaker protein-protein interactions (bipartite binding). The EDEM1 mannosidase-like domain (MLD) alone retains both thiol-dependent binding and glycan-trimming activity. Two intrinsically disordered regions (IDRs), one N-terminal and one C-terminal, are both required for ERdj5 binding.","method":"Co-immunoprecipitation under reducing/non-reducing conditions; Cys-to-Ala point mutation in client; MLD domain construct; IDR deletion mutants","journal":"The Journal of Biological Chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple domain mutants and redox manipulation, single lab","pmids":["30021839"],"is_preprint":false},{"year":2020,"finding":"EDEM1 is found in auto-regulatory complexes with ERAD components (identified by mass spectrometry). The N-terminal disordered region of EDEM1 mediates protein-protein interaction with misfolded proteins; deletion of this domain significantly impairs their degradation. When proteasomal activity is severely impaired, EDEM1 overexpression can still promote degradation by promoting aggregate formation, which is then cleared by autophagy (ER-phagy as a back-up).","method":"Mass spectrometry; co-immunoprecipitation; EDEM1 domain deletion mutants; proteasome inhibition with MG132; autophagy assays","journal":"International Journal of Molecular Sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MS-confirmed complex plus functional domain analysis, single lab","pmids":["32423001"],"is_preprint":false},{"year":2021,"finding":"EDEM1 and EDEM3 are responsible for the second step of N-glycan mannose trimming in ERAD (from M8B to M7, M6, M5, exposing the alpha-1,6-linked mannosyl residue). Purified EDEM3 alone converted pyridylamine-labeled M8B to M7A, M7C, M6, and M5; purified EDEM1 showed similar but weaker activity (less M6/M5). Both efficiently trimmed M8B from a glycoprotein substrate.","method":"In vitro mannosidase assay with purified EDEM1 and EDEM3 proteins; pyridylamine-labeled M8B oligosaccharide and glycoprotein substrates; HPLC N-glycan profiling","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1 / Moderate — reconstituted in vitro enzymatic assay with purified proteins establishing catalytic step, single lab","pmids":["34698634"],"is_preprint":false},{"year":2021,"finding":"EDEM1 physically interacts with amyloid precursor protein (APP) and promotes APP degradation via ERAD retrotranslocation to the cytosol. EDEM1 overexpression reduced APP cellular levels and decreased Abeta40/Abeta42 secretion; EDEM1 knockdown increased APP levels.","method":"Co-immunoprecipitation; overexpression and siRNA knockdown; pulse-chase assays; Abeta ELISA","journal":"International Journal of Molecular Sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain-of-function, loss-of-function, and direct binding assay, single lab","pmids":["35008544"],"is_preprint":false},{"year":2023,"finding":"EDEM1 physically associates with EGFR and enhances EGFR degradation via ERAD. EGFR and thrombospondin-1 (TSP1) were identified as endogenous EDEM1 substrate proteins; their protein maturation status and cellular localization were markedly affected by EDEM1 knockdown.","method":"siRNA knockdown of EDEM1; co-immunoprecipitation; pulse-chase and protein stability assays; immunofluorescence","journal":"International Journal of Molecular Sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss-of-function with direct binding assay for physiological substrates, single lab","pmids":["37569550"],"is_preprint":false},{"year":2024,"finding":"EDEM1 is itself turned over by both ERAD and autophagy. ERAD-dependent degradation of EDEM1 involves the SEL1L/HRD1 complex, YOD1, XTP3-B, ERdj3, VIMP, BAG6, and JB12, but not OS9, and occurs in both mannose-trimming-dependent and -independent manners.","method":"Sibling knockdowns of ERAD components; proteasome and autophagy inhibitors; pulse-chase assays","journal":"Genes to Cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — systematic genetic knockdown of pathway components, single lab","pmids":["38682256"],"is_preprint":false},{"year":2018,"finding":"Silencing of EDEM1 increased bioavailability of ATF6 for Golgi export and cleavage upon ER stress, by stabilizing the natively unstable ATF6 protein. A somatic cancer variant of EDEM1 (N198I) altered ATF6 signaling. This places EDEM1 as a negative regulator of ATF6 stability/activity.","method":"siRNA screening; ATF6 export and cleavage assays; overexpression of EDEM1 variants","journal":"The FEBS Journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — phenotypic siRNA screen with mechanistic follow-up, single lab","pmids":["30281916"],"is_preprint":false},{"year":2023,"finding":"EDEM1 inhibits the IRE1/JNK/c-Jun signaling pathway, leading to increased insulin mRNA levels. EDEM1 overexpression in INS-1E beta cells and human islets increased insulin secretion upon glucose stimulation; EDEM1 modulates UPR via both IRE1/XBP1s and IRE1/JNK/c-Jun cascades.","method":"Overexpression in INS-1E cells and human islets; western blotting of IRE1/JNK/c-Jun pathway; qRT-PCR for insulin mRNA; glucose-stimulated insulin secretion assay; in vivo diabetic rat model","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain-of-function in multiple cell types with pathway readouts, single lab","pmids":["37822496"],"is_preprint":false},{"year":2025,"finding":"Cryo-EM structures of the EDEM:PDI heterodimer (from Chaetomium thermophilum orthologs) were solved, with and without an alpha1-antitrypsin NHK client. The EDEM catalytic domain nests within the PDI arc; the client (A1AT-NHK) binds EDEM's C-terminal flexible domains. A disulfide bond forms between A1AT-NHK and an exposed Cys in the EDEM PAD domain. Redox chemistry between EDEM and PDI generates oxidized (demannosylation-competent) EDEM and reduced PDI, priming PDI to act as the ERAD reductase facilitating client retrotranslocation.","method":"Cryo-EM structure determination; mass spectrometry (disulfide bond identification); non-reducing SDS-PAGE after co-transfection; in vitro redox assays","journal":"bioRxiv (preprint)","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — cryo-EM structures with MS validation and functional biochemistry, single lab, preprint not yet peer-reviewed","pmids":["bio_10.1101_2025.01.29.635535"],"is_preprint":true}],"current_model":"EDEM1 is an ER-resident mannosidase-like protein that extracts terminally misfolded glycoproteins (and some nonglycosylated proteins) from the calnexin chaperone cycle via its N-terminal intrinsically disordered region and redox-sensitive thiol interactions, then targets them to the SEL1L/HRD1 dislocation complex for retrotranslocation and proteasomal degradation; it possesses intrinsic alpha-1,2-mannosidase catalytic activity (trimming the C branch of N-glycans, enhanced on unfolded substrates and by association with TXNDC11/PDI), prevents covalent aggregation of ERAD substrates, exists as both soluble and membrane-anchored isoforms that associate differentially with ERdj5 or SEL1L, traffics through non-COPII vesicles (EDEMosomes) out of the ER, and is itself turned over by selective autophagy (after cytosolic PNGase-mediated deglycosylation and p62/NBR1-dependent targeting) and by the ERAD/SEL1L-HRD1 pathway."},"narrative":{"mechanistic_narrative":"EDEM1 is an endoplasmic reticulum-resident mannosidase-like protein that commits terminally misfolded glycoproteins to ER-associated degradation (ERAD) by extracting them from the calnexin folding cycle and routing them to the dislocation machinery [PMID:12610306, PMID:12610305]. It physically associates with calnexin and acts as an acceptor that releases folding-incompetent substrates, accelerating their degradation while sparing productively folding clients [PMID:12610306, PMID:12610305]. EDEM1 possesses intrinsic alpha-1,2-mannosidase activity, trimming the C branch of N-glycans and catalyzing the second mannose-trimming step (M8B to M7/M6/M5) that exposes the alpha-1,6-mannose recognized by downstream ERAD lectins; this activity is modest on native substrates but strongly enhanced on unfolded glycoproteins and by association with oxidoreductases such as TXNDC11 [PMID:20065073, PMID:30374462, PMID:34698634]. Substrate recognition is genetically separable from catalysis: EDEM1 binds nonnative proteins through its N-terminal intrinsically disordered region and through hydrophobicity- and thiol-dependent contacts, independently of glycan trimming, allowing it to engage both glycosylated and nonglycosylated clients and to prevent their covalent disulfide-linked aggregation during release from calnexin [PMID:16987498, PMID:19524542, PMID:23233672, PMID:22905195, PMID:25655076, PMID:30021839]. EDEM1 then delivers clients to the SEL1L/HRD1 dislocation and ubiquitination complex, positioning it upstream of the mannose-dependent lectins XTP3-B and OS9 [PMID:19524542, PMID:21062743, PMID:21917589]. Two isoforms arising from inefficient signal-sequence cleavage partition this activity: a soluble form associates with the oxidoreductase ERdj5 to handle soluble substrates and a type-II membrane form associates with SEL1L for membrane substrates [PMID:21632540]. EDEM1 promotes retrotranslocation of physiological and toxin clients including ricin A-chain, APP, and EGFR, and is itself turned over by selective autophagy following PNGase deglycosylation and p62/NBR1 targeting, as well as by the SEL1L/HRD1 ERAD pathway [PMID:16452630, PMID:21388347, PMID:24664425, PMID:35008544, PMID:37569550, PMID:38682256]. Beyond degradation, EDEM1 modulates the unfolded protein response by negatively regulating ATF6 stability and by tuning IRE1 signaling [PMID:30281916, PMID:37822496].","teleology":[{"year":2003,"claim":"Established EDEM1's core role by showing it extracts terminally misfolded glycoproteins from the calnexin cycle, answering how doomed clients are diverted from futile folding into degradation.","evidence":"Overexpression/knockdown with pulse-chase of misfolded glycoprotein fate and reciprocal co-IP mapping calnexin binding to the transmembrane region","pmids":["12610306","12610305"],"confidence":"High","gaps":["Did not establish the biochemical signal distinguishing misfolded from folding-competent clients","Catalytic activity not yet demonstrated"]},{"year":2004,"claim":"Resolved that EDEM1 also exists as a soluble ER-lumenal species and defined a stress-regulated paralog EDEM2, clarifying that EDEM function is not restricted to a membrane-anchored form.","evidence":"Subcellular fractionation, pulse-chase ERAD assays, and XBP1 reporter assays","pmids":["15579471"],"confidence":"Medium","gaps":["Relationship between soluble and membrane forms unresolved","Functional distinction between EDEM1 and EDEM2 substrates unaddressed"]},{"year":2006,"claim":"Dissected two separable EDEM1 activities by catalytic-site mutagenesis, showing de-mannosylation and anti-aggregation chaperone function are mechanistically distinct.","evidence":"Site-directed mutagenesis of the conserved alpha-1,2-mannosidase catalytic residue (E220Q) with N-glycan analysis and non-reducing SDS-PAGE; comparison against control ER proteins","pmids":["16987498","16629899"],"confidence":"High","gaps":["Direct enzymatic activity of purified protein not shown","Molecular basis of anti-aggregation function undefined"]},{"year":2007,"claim":"Revealed a non-canonical ER exit route by localizing endogenous EDEM1 to non-COPII ~150 nm vesicles (EDEMosomes) shared with Derlin-2 and misfolded substrate.","evidence":"Quantitative immunogold electron microscopy with serial sections and subcellular fractionation","pmids":["17360537"],"confidence":"High","gaps":["Machinery generating these vesicles unidentified","Destination and functional purpose of vesicular EDEM1 unresolved"]},{"year":2009,"claim":"Decoupled substrate recognition from glycan chemistry, showing EDEM1 binds nonnative proteins glycan-independently while its mannosidase-like domain targets clients to SEL1L.","evidence":"Co-IP under kifunensine, mannosidase-domain mutants, and SEL1L pulldowns; extended by an opsin chaperone study and the first demonstration of basal autophagic turnover of EDEM1","pmids":["19524542","19934218","19266160"],"confidence":"High","gaps":["The protein determinant recognized by EDEM1 not yet defined","How recognition and SEL1L handoff are coordinated unclear"]},{"year":2011,"claim":"Placed EDEM1 upstream of mannose-dependent lectins and showed its overexpression can bypass the ER mannosidase I requirement, refining the ERAD step order.","evidence":"ERManI knockdown with EDEM1 overexpression, deletion mutants lacking the carbohydrate-recognition domain, and colocalization at the pericentriolar ERQC; isoform analysis defining soluble-ERdj5 vs membrane-SEL1L partitioning","pmids":["21062743","21917589","21632540"],"confidence":"Medium","gaps":["Quantitative contribution of mannose-dependent vs -independent routing in vivo unresolved","Regulation of isoform ratio not defined"]},{"year":2015,"claim":"Generalized EDEM1's client repertoire to nonglycosylated proteins and toxin chains, and identified hydrophobicity and a disordered N-terminal region as the recognition basis.","evidence":"Domain deletion and substrate-hydrophobicity mutagenesis with co-IP and ERAD/retrotranslocation assays across multiple substrates (NHK, H2a, ricin A-chain, BACE457, tyrosinase)","pmids":["23233672","22905195","25655076","21388347"],"confidence":"Medium","gaps":["Structural details of disordered-region engagement unresolved","Whether the same surface binds glycosylated and nonglycosylated clients unclear"]},{"year":2018,"claim":"Demonstrated intrinsic mannosidase activity directly in vitro and showed it is substrate-state dependent and enhanced by oxidoreductase partners, and that client capture involves redox-sensitive thiol contacts.","evidence":"In vitro mannosidase assays with purified EDEM1/2 on native vs denatured substrates, co-IP with TXNDC11; reducing/non-reducing co-IP with client Cys256 mutants and IDR/MLD constructs","pmids":["30374462","30021839"],"confidence":"High","gaps":["Stoichiometry and dynamics of oxidoreductase coupling in cells unresolved","How redox state gates substrate release undefined"]},{"year":2021,"claim":"Defined the precise enzymatic step EDEM1 catalyzes (M8B to M7/M6/M5) using reconstituted purified enzyme, situating it alongside EDEM3 in N-glycan trimming.","evidence":"In vitro mannosidase assay with purified EDEM1 and EDEM3 on pyridylamine-labeled M8B and glycoprotein substrates with HPLC profiling","pmids":["34698634"],"confidence":"High","gaps":["Functional division of labor between EDEM1 and EDEM3 in vivo unresolved","Kinetic parameters on physiological clients undefined"]},{"year":2023,"claim":"Extended EDEM1 to physiological and disease-relevant clients (APP, EGFR, TSP1) and to UPR regulation, showing roles beyond bulk misfolded-protein clearance.","evidence":"Knockdown/overexpression with co-IP, pulse-chase, Abeta ELISA, and maturation/localization readouts; ATF6 export/cleavage assays and IRE1/JNK/c-Jun pathway analysis with insulin secretion in beta cells and islets","pmids":["35008544","37569550","30281916","37822496"],"confidence":"Medium","gaps":["Direct vs indirect effects on UPR transducers not fully separated","Physiological significance of individual substrate effects in tissue not established"]},{"year":2024,"claim":"Mapped how EDEM1 itself is degraded, showing dual ERAD (SEL1L/HRD1, YOD1, XTP3-B, ERdj3, VIMP, BAG6, JB12) and selective autophagy routes, establishing autoregulation of the ERAD machinery.","evidence":"Sibling knockdowns of ERAD components, proteasome and autophagy inhibitors, pulse-chase; PNGase-dependent p62/NBR1 routing imaging and autoregulatory complex mass spectrometry","pmids":["38682256","24664425","32423001"],"confidence":"Medium","gaps":["Trigger selecting ERAD vs autophagy for EDEM1 turnover unresolved","Physiological consequences of dysregulated EDEM1 levels undefined"]},{"year":2025,"claim":"Provided the first structural model of EDEM client engagement, showing the EDEM catalytic domain nesting in a PDI arc with disulfide-mediated client capture and a redox relay priming PDI as the ERAD reductase.","evidence":"Cryo-EM of EDEM:PDI heterodimer with/without A1AT-NHK client (thermophilic orthologs), mass spectrometry of disulfide bonds, and in vitro redox assays (preprint)","pmids":["bio_10.1101_2025.01.29.635535"],"confidence":"Medium","gaps":["Awaits peer review and confirmation in human EDEM1","How redox cycling couples to retrotranslocation in the full dislocon unresolved"]},{"year":null,"claim":"It remains unresolved how EDEM1 integrates substrate recognition, catalytic trimming, redox chemistry, and isoform/vesicle partitioning into a single regulated decision to commit a client to degradation in vivo.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No quantitative model linking mannose-dependent and -independent routing","Physiological regulation of EDEMosome formation and EDEM1 turnover unknown","No high-resolution human EDEM1 structure with client"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140098","term_label":"catalytic activity, acting on RNA","supporting_discovery_ids":[9,18,21]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[9,18,21]},{"term_id":"GO:0044183","term_label":"protein folding chaperone","supporting_discovery_ids":[3,4]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[6,14,19]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[2,5,12]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[5]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,1,6,24]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[25,26]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[8,16,20]}],"complexes":["SEL1L/HRD1 ERAD dislocation complex","EDEM:PDI heterodimer"],"partners":["CANX","SEL1L","DNAJC10","TXNDC11","SQSTM1","NBR1","PDIA"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q92611","full_name":"ER degradation-enhancing alpha-mannosidase-like protein 1","aliases":[],"length_aa":657,"mass_kda":73.8,"function":"Extracts misfolded glycoproteins, but not glycoproteins undergoing productive folding, from the calnexin cycle. It is directly involved in endoplasmic reticulum-associated degradation (ERAD) and targets misfolded glycoproteins for degradation in an N-glycan-independent manner, probably by forming a complex with SEL1L. 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DERL2","url":"https://www.omim.org/entry/610304"},{"mim_id":"610302","title":"ENDOPLASMIC RETICULUM DEGRADATION-ENHANCING ALPHA-MANNOSIDASE-LIKE PROTEIN 2; EDEM2","url":"https://www.omim.org/entry/610302"},{"mim_id":"607673","title":"ENDOPLASMIC RETICULUM DEGRADATION-ENHANCING ALPHA-MANNOSIDASE-LIKE PROTEIN 1; EDEM1","url":"https://www.omim.org/entry/607673"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Endoplasmic reticulum","reliability":"Enhanced"},{"location":"Aggresome","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"lymphoid 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EDEM overexpression caused faster release of folding-incompetent proteins from calnexin and earlier onset of degradation; EDEM down-regulation prolonged folding attempts and delayed ERAD.\",\n      \"method\": \"Overexpression and down-regulation of EDEM combined with pulse-chase analysis of misfolded glycoprotein fate and calnexin association\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — two independent labs (PMIDs 12610306 and 12610305) published simultaneously with complementary functional evidence (overexpression, knockdown, co-immunoprecipitation, pulse-chase)\",\n      \"pmids\": [\"12610306\", \"12610305\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"EDEM interacts physically with calnexin through its transmembrane region (not with calreticulin). Both binding of substrates to calnexin and their release from calnexin are required for ERAD; EDEM overexpression promoted release of terminally misfolded proteins from calnexin, functioning as an acceptor of calnexin substrates.\",\n      \"method\": \"Co-immunoprecipitation; overexpression studies with pulse-chase analysis; domain deletion mapping (transmembrane region)\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP identifying binding partner and domain, replicated finding across two simultaneous independent publications\",\n      \"pmids\": [\"12610305\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"EDEM is a soluble protein of the ER lumen (in HEK293 cells), not exclusively an ER membrane protein; it functions as a mannosidase-like chaperone. A second homolog EDEM2 was identified that is also a stress-regulated mannosidase-like protein operating in the ER lumen and accelerates ERAD of terminally misfolded glycoproteins by facilitating their extraction from the calnexin cycle; EDEM2 transcriptional up-regulation depends on the ER stress-activated transcription factor XBP1.\",\n      \"method\": \"Subcellular fractionation; overexpression and pulse-chase ERAD assays; reporter assays for XBP1-dependent transcription\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — fractionation establishing soluble localization plus functional ERAD assay, single lab\",\n      \"pmids\": [\"15579471\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"EDEM1 overexpression accelerates de-mannosylation of terminally misfolded glycoproteins (Man9 or Man5 N-glycans) and inhibits formation of covalent (disulfide-linked) aggregates upon their release from calnexin. Substitution of one conserved catalytic residue in the alpha-1,2-mannosidase active site (E220Q equivalent) fully blocked accelerated de-mannosylation but did not affect the anti-aggregation (chaperone) function of EDEM1, indicating two separable activities.\",\n      \"method\": \"Pulse-chase with N-glycan analysis; site-directed mutagenesis of conserved catalytic residue; non-reducing SDS-PAGE to detect aggregates\",\n      \"journal\": \"Biochemical and Biophysical Research Communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — active-site mutagenesis dissecting two activities (mannosidase vs. chaperone) with multiple orthogonal methods, single lab\",\n      \"pmids\": [\"16987498\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"EDEM prevents aberrant covalent dimer formation of misfolded alpha1-antitrypsin NHK, maintaining its retrotranslocation competence. EDEM overexpression selectively prevented accumulation of covalent NHK dimers; this anti-aggregation effect was specific to EDEM and not observed with calnexin or H+/K+-ATPase beta subunit overexpression.\",\n      \"method\": \"Overexpression of EDEM and control ER membrane proteins; non-reducing SDS-PAGE; pulse-chase degradation assays\",\n      \"journal\": \"Genes to Cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean functional comparison with controls, single lab\",\n      \"pmids\": [\"16629899\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Endogenous EDEM1 is sequestered in ~150 nm vesicles that bud from rough ER cisternae outside of transitional ER exit sites, lacking a recognizable COPII coat (~87% of immunogold label on vesicles). These vesicles also contain Derlin-2 and misfolded alpha-1-antitrypsin (NHK), indicating a non-canonical ER exit pathway for quality control components and substrates.\",\n      \"method\": \"High-resolution immunogold electron microscopy and serial section analysis; subcellular fractionation\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the USA\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — high-resolution structural imaging with quantitative immunogold labeling demonstrating non-COPII vesicle pathway, single lab with rigorous controls\",\n      \"pmids\": [\"17360537\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"EDEM1 specifically binds nonnative proteins in a glycan-independent manner. Inhibition of mannosidase activity with kifunensine or mutation of the mannosidase-like domain had no effect on substrate binding but diminished EDEM1 association with the ER membrane adaptor protein SEL1L. EDEM1 thus uses its mannosidase-like domain to target aberrant proteins to the SEL1L-containing dislocation/ubiquitination complex.\",\n      \"method\": \"Co-immunoprecipitation under glycan-inhibited conditions (kifunensine); mannosidase domain mutants; pulldown assays with SEL1L\",\n      \"journal\": \"Molecular Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP, domain mutagenesis, pharmacological inhibition as orthogonal methods, single lab\",\n      \"pmids\": [\"19524542\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"EDEM1 functions as a chaperone for rod opsin: it promotes degradation of misfolded P23H rod opsin via ERAD and decreases its aggregation into inclusions. shRNA knockdown of EDEM1 increased P23H rod opsin levels and aggregation. Unexpectedly, EDEM1 binding to rod opsin was independent of mannose trimming, and EDEM1 also promoted cell-surface expression of mutant rod opsin.\",\n      \"method\": \"shRNA knockdown; overexpression; co-immunoprecipitation; pulse-chase assays; immunofluorescence; endoglycosidase H sensitivity assays\",\n      \"journal\": \"Journal of Cell Science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function (shRNA) and gain-of-function with multiple readouts, single lab\",\n      \"pmids\": [\"19934218\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Endogenous EDEM1 is degraded by basal autophagy (not by the proteasome under non-starved conditions). EDEM1 was detected in autophagosomes biochemically (LC3 immuno-purification) and by immunocytochemistry. Inhibition of the lysosome-autophagy pathway or knockdown of ATG genes stabilized EDEM1.\",\n      \"method\": \"Autophagosome immunopurification; immunocytochemistry; pharmacological inhibitors (vinblastine, pepstatin A/E64d, 3-methyladenine); siRNA knockdown of ATGs\",\n      \"journal\": \"Cellular and Molecular Life Sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal approaches (biochemical fractionation, pharmacology, siRNA), single lab\",\n      \"pmids\": [\"19266160\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"EDEM1 overexpression specifically accelerates trimming of alpha-1,2-linked mannose from the C branch of N-glycans, producing Glc1Man8GlcNAc2 isomer C and increasing Man7GlcNAc2 isomer A. The EDEM1 E220Q catalytic mutant did not produce this isomer, confirming alpha-1,2-mannosidase catalytic activity on the C branch. However, ERAD enhancement by wild-type and E220Q EDEM1 was equivalent, attributed to inhibition of aberrant NHK dimer formation.\",\n      \"method\": \"Radiolabeled N-glycan structural analysis (3H-mannose pulse-chase); site-directed mutagenesis (E220Q); stable cell line overexpression\",\n      \"journal\": \"Glycobiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vivo glycan structural analysis with catalytic mutant, rigorous N-glycan mapping, single lab\",\n      \"pmids\": [\"20065073\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Mannose trimming is not required for EDEM1 binding to an ERAD substrate glycoprotein (binding persists with kifunensine treatment or ERManI knockdown). In contrast, substrate association with XTP3-B and E3 ubiquitin ligases HRD1 and SCF(Fbs2) required mannose trimming, placing EDEM1 upstream of these factors in the ERAD pathway.\",\n      \"method\": \"Co-immunoprecipitation under mannosidase inhibitor (kifunensine) treatment; ERManI siRNA knockdown; colocalization by immunofluorescence at the ERQC\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological and genetic inhibition with co-IP and colocalization, single lab\",\n      \"pmids\": [\"21062743\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"EDEM1 overexpression or its up-regulation via IRE1 (unfolded protein response) overrides the mannose-trimming requirement for ERAD, rendering ER mannosidase I dispensable. An EDEM1 deletion mutant lacking most of the carbohydrate-recognition domain still accelerated ERAD and delivered substrate to XTP3-B and OS9. Upon proteasomal inhibition, EDEM1 concentrated with ERAD substrate in the pericentriolar ERQC compartment containing OS9.\",\n      \"method\": \"ERManI knockdown combined with EDEM1 overexpression; EDEM1 deletion mutants; co-immunoprecipitation; immunofluorescence colocalization at ERQC\",\n      \"journal\": \"Molecular Biology of the Cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain deletion analysis and epistasis experiments, single lab\",\n      \"pmids\": [\"21917589\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Signal sequence cleavage of EDEM1 is slow and inefficient, producing two isoforms: a soluble form and a type-II membrane form (when signal sequence is uncleaved, creating an N-terminal transmembrane segment). The soluble form efficiently associates with the oxidoreductase ERdj5 and accelerates turnover of soluble ERAD substrates, while the membrane form efficiently associates with SEL1L and accelerates turnover of membrane-associated ERAD substrates.\",\n      \"method\": \"Signal sequence cleavage analysis; glycosylation site mapping; co-immunoprecipitation with ERdj5 and SEL1L; pulse-chase ERAD assays with isoform-specific constructs\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical isoform characterization with functional ERAD assays, single lab\",\n      \"pmids\": [\"21632540\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"EDEM1 participates in a shared ERAD pathway for both glycosylated and nonglycosylated proteins. Nonglycosylated ERAD substrates (including nonglycosylated H2a, NS-1κ light chain, truncated Igγ heavy chain) co-immunoprecipitate with EDEM1, and require EDEM1 for their degradation. EDEM1 associates with nonglycosylated proteins through a region outside its mannosidase-like domain.\",\n      \"method\": \"Co-immunoprecipitation; siRNA knockdown of EDEM1; pulse-chase degradation assays; domain deletion analysis\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple substrates, domain mapping, loss-of-function, single lab\",\n      \"pmids\": [\"23233672\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"The intrinsically disordered (ID) N-terminal region of EDEM1 (residues ~40–119) mediates binding to both glycosylated and non-glycosylated misfolded proteins. Deletion of this ID region abolished co-immunoprecipitation with misfolded tyrosinase (glycosylated and non-glycosylated mutants), while the intact mannosidase-like domain was not required for substrate binding. EDEM1 overexpression enhanced degradation of wild-type and misfolded tyrosinase.\",\n      \"method\": \"Co-immunoprecipitation with EDEM1 deletion mutants; homology modeling of mannosidase domain; pulse-chase degradation assays\",\n      \"journal\": \"PLOS ONE\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain deletion mutants with co-IP and functional assays, single lab\",\n      \"pmids\": [\"22905195\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"EDEM1 promotes retrotranslocation of ricin A-chain (RTA) from the ER to the cytosol. EDEM1 overexpression increased RTA retrotranslocation (demonstrated with kifunensine to block competition from misfolded proteins), while RNAi-mediated knockdown decreased RTA retrotranslocation. Co-immunoprecipitation showed that ricin interacts with EDEM1 and with Sec61alpha.\",\n      \"method\": \"Overexpression and siRNA knockdown of EDEM1; cytosol/ER fractionation to measure retrotranslocation; co-immunoprecipitation\",\n      \"journal\": \"Molecular Biology of the Cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function and gain-of-function with biochemical fractionation, single lab (replication of PMID 16452630 results)\",\n      \"pmids\": [\"16452630\", \"21388347\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"EDEM1 is degraded by selective autophagy. It colocalizes with selective autophagy cargo receptors p62/SQSTM1, NBR1, and Alfy, and is engulfed by autophagic isolation membranes. p62/SQSTM1 and NBR1 knockdown blocked EDEM1 routing to autophagosomes. p62/SQSTM1 interacts only with deglycosylated (and ubiquitinated) EDEM1; deglycosylation by cytosolic peptide N-glycanase (PNGase) is a prerequisite for p62/SQSTM1 interaction and aggregate formation.\",\n      \"method\": \"Immunofluorescence colocalization; siRNA knockdown of cargo receptors; PNGase inhibitors; ubiquitination assays\",\n      \"journal\": \"Histochemistry and Cell Biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal approaches (siRNA, inhibitors, imaging), single lab\",\n      \"pmids\": [\"24664425\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Recognition of ERAD substrates by EDEM1 is determined by the hydrophobicity of protein determinants. Mutations increasing or decreasing hydrophobicity in misfolded substrates (ricin A-chain, BACE457) correspondingly altered their interaction with EDEM1. EDEM1 can bind hydrophobic transmembrane regions of misfolded ERAD substrates, and this binding does not require substrate glycosylation.\",\n      \"method\": \"Site-directed mutagenesis to alter substrate hydrophobicity; co-immunoprecipitation; ERAD assays\",\n      \"journal\": \"BMC Cell Biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mutagenesis-driven substrate binding analysis, single lab\",\n      \"pmids\": [\"25655076\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"EDEM1 and EDEM2 possess intrinsic mannosidase activity in vitro, demonstrated directly. Their activity on free N-glycans and intact glycoproteins is modest, but is significantly enhanced on denatured (unfolded) glycoproteins. EDEM1/2 associate with oxidoreductases including TXNDC11, which enhances their mannosidase activity on glycoproteins but not on free N-glycans.\",\n      \"method\": \"In vitro mannosidase assay with purified proteins; denatured vs. native substrate comparisons; co-immunoprecipitation with oxidoreductases\",\n      \"journal\": \"Communications Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — direct in vitro enzymatic assay establishing catalytic activity and substrate-state dependence, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"30374462\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"EDEM1 binds ERAD clients (Z and NHK alpha1-antitrypsin) through a thiol-dependent (redox-sensitive) interaction via Cys256 of the client, as well as through weaker protein-protein interactions (bipartite binding). The EDEM1 mannosidase-like domain (MLD) alone retains both thiol-dependent binding and glycan-trimming activity. Two intrinsically disordered regions (IDRs), one N-terminal and one C-terminal, are both required for ERdj5 binding.\",\n      \"method\": \"Co-immunoprecipitation under reducing/non-reducing conditions; Cys-to-Ala point mutation in client; MLD domain construct; IDR deletion mutants\",\n      \"journal\": \"The Journal of Biological Chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple domain mutants and redox manipulation, single lab\",\n      \"pmids\": [\"30021839\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"EDEM1 is found in auto-regulatory complexes with ERAD components (identified by mass spectrometry). The N-terminal disordered region of EDEM1 mediates protein-protein interaction with misfolded proteins; deletion of this domain significantly impairs their degradation. When proteasomal activity is severely impaired, EDEM1 overexpression can still promote degradation by promoting aggregate formation, which is then cleared by autophagy (ER-phagy as a back-up).\",\n      \"method\": \"Mass spectrometry; co-immunoprecipitation; EDEM1 domain deletion mutants; proteasome inhibition with MG132; autophagy assays\",\n      \"journal\": \"International Journal of Molecular Sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MS-confirmed complex plus functional domain analysis, single lab\",\n      \"pmids\": [\"32423001\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"EDEM1 and EDEM3 are responsible for the second step of N-glycan mannose trimming in ERAD (from M8B to M7, M6, M5, exposing the alpha-1,6-linked mannosyl residue). Purified EDEM3 alone converted pyridylamine-labeled M8B to M7A, M7C, M6, and M5; purified EDEM1 showed similar but weaker activity (less M6/M5). Both efficiently trimmed M8B from a glycoprotein substrate.\",\n      \"method\": \"In vitro mannosidase assay with purified EDEM1 and EDEM3 proteins; pyridylamine-labeled M8B oligosaccharide and glycoprotein substrates; HPLC N-glycan profiling\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — reconstituted in vitro enzymatic assay with purified proteins establishing catalytic step, single lab\",\n      \"pmids\": [\"34698634\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"EDEM1 physically interacts with amyloid precursor protein (APP) and promotes APP degradation via ERAD retrotranslocation to the cytosol. EDEM1 overexpression reduced APP cellular levels and decreased Abeta40/Abeta42 secretion; EDEM1 knockdown increased APP levels.\",\n      \"method\": \"Co-immunoprecipitation; overexpression and siRNA knockdown; pulse-chase assays; Abeta ELISA\",\n      \"journal\": \"International Journal of Molecular Sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain-of-function, loss-of-function, and direct binding assay, single lab\",\n      \"pmids\": [\"35008544\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EDEM1 physically associates with EGFR and enhances EGFR degradation via ERAD. EGFR and thrombospondin-1 (TSP1) were identified as endogenous EDEM1 substrate proteins; their protein maturation status and cellular localization were markedly affected by EDEM1 knockdown.\",\n      \"method\": \"siRNA knockdown of EDEM1; co-immunoprecipitation; pulse-chase and protein stability assays; immunofluorescence\",\n      \"journal\": \"International Journal of Molecular Sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function with direct binding assay for physiological substrates, single lab\",\n      \"pmids\": [\"37569550\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"EDEM1 is itself turned over by both ERAD and autophagy. ERAD-dependent degradation of EDEM1 involves the SEL1L/HRD1 complex, YOD1, XTP3-B, ERdj3, VIMP, BAG6, and JB12, but not OS9, and occurs in both mannose-trimming-dependent and -independent manners.\",\n      \"method\": \"Sibling knockdowns of ERAD components; proteasome and autophagy inhibitors; pulse-chase assays\",\n      \"journal\": \"Genes to Cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic genetic knockdown of pathway components, single lab\",\n      \"pmids\": [\"38682256\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Silencing of EDEM1 increased bioavailability of ATF6 for Golgi export and cleavage upon ER stress, by stabilizing the natively unstable ATF6 protein. A somatic cancer variant of EDEM1 (N198I) altered ATF6 signaling. This places EDEM1 as a negative regulator of ATF6 stability/activity.\",\n      \"method\": \"siRNA screening; ATF6 export and cleavage assays; overexpression of EDEM1 variants\",\n      \"journal\": \"The FEBS Journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phenotypic siRNA screen with mechanistic follow-up, single lab\",\n      \"pmids\": [\"30281916\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EDEM1 inhibits the IRE1/JNK/c-Jun signaling pathway, leading to increased insulin mRNA levels. EDEM1 overexpression in INS-1E beta cells and human islets increased insulin secretion upon glucose stimulation; EDEM1 modulates UPR via both IRE1/XBP1s and IRE1/JNK/c-Jun cascades.\",\n      \"method\": \"Overexpression in INS-1E cells and human islets; western blotting of IRE1/JNK/c-Jun pathway; qRT-PCR for insulin mRNA; glucose-stimulated insulin secretion assay; in vivo diabetic rat model\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain-of-function in multiple cell types with pathway readouts, single lab\",\n      \"pmids\": [\"37822496\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Cryo-EM structures of the EDEM:PDI heterodimer (from Chaetomium thermophilum orthologs) were solved, with and without an alpha1-antitrypsin NHK client. The EDEM catalytic domain nests within the PDI arc; the client (A1AT-NHK) binds EDEM's C-terminal flexible domains. A disulfide bond forms between A1AT-NHK and an exposed Cys in the EDEM PAD domain. Redox chemistry between EDEM and PDI generates oxidized (demannosylation-competent) EDEM and reduced PDI, priming PDI to act as the ERAD reductase facilitating client retrotranslocation.\",\n      \"method\": \"Cryo-EM structure determination; mass spectrometry (disulfide bond identification); non-reducing SDS-PAGE after co-transfection; in vitro redox assays\",\n      \"journal\": \"bioRxiv (preprint)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — cryo-EM structures with MS validation and functional biochemistry, single lab, preprint not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.01.29.635535\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"EDEM1 is an ER-resident mannosidase-like protein that extracts terminally misfolded glycoproteins (and some nonglycosylated proteins) from the calnexin chaperone cycle via its N-terminal intrinsically disordered region and redox-sensitive thiol interactions, then targets them to the SEL1L/HRD1 dislocation complex for retrotranslocation and proteasomal degradation; it possesses intrinsic alpha-1,2-mannosidase catalytic activity (trimming the C branch of N-glycans, enhanced on unfolded substrates and by association with TXNDC11/PDI), prevents covalent aggregation of ERAD substrates, exists as both soluble and membrane-anchored isoforms that associate differentially with ERdj5 or SEL1L, traffics through non-COPII vesicles (EDEMosomes) out of the ER, and is itself turned over by selective autophagy (after cytosolic PNGase-mediated deglycosylation and p62/NBR1-dependent targeting) and by the ERAD/SEL1L-HRD1 pathway.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EDEM1 is an endoplasmic reticulum-resident mannosidase-like protein that commits terminally misfolded glycoproteins to ER-associated degradation (ERAD) by extracting them from the calnexin folding cycle and routing them to the dislocation machinery [#0, #1]. It physically associates with calnexin and acts as an acceptor that releases folding-incompetent substrates, accelerating their degradation while sparing productively folding clients [#0, #1]. EDEM1 possesses intrinsic alpha-1,2-mannosidase activity, trimming the C branch of N-glycans and catalyzing the second mannose-trimming step (M8B to M7/M6/M5) that exposes the alpha-1,6-mannose recognized by downstream ERAD lectins; this activity is modest on native substrates but strongly enhanced on unfolded glycoproteins and by association with oxidoreductases such as TXNDC11 [#9, #18, #21]. Substrate recognition is genetically separable from catalysis: EDEM1 binds nonnative proteins through its N-terminal intrinsically disordered region and through hydrophobicity- and thiol-dependent contacts, independently of glycan trimming, allowing it to engage both glycosylated and nonglycosylated clients and to prevent their covalent disulfide-linked aggregation during release from calnexin [#3, #6, #13, #14, #17, #19]. EDEM1 then delivers clients to the SEL1L/HRD1 dislocation and ubiquitination complex, positioning it upstream of the mannose-dependent lectins XTP3-B and OS9 [#6, #10, #11]. Two isoforms arising from inefficient signal-sequence cleavage partition this activity: a soluble form associates with the oxidoreductase ERdj5 to handle soluble substrates and a type-II membrane form associates with SEL1L for membrane substrates [#12]. EDEM1 promotes retrotranslocation of physiological and toxin clients including ricin A-chain, APP, and EGFR, and is itself turned over by selective autophagy following PNGase deglycosylation and p62/NBR1 targeting, as well as by the SEL1L/HRD1 ERAD pathway [#15, #16, #22, #23, #24]. Beyond degradation, EDEM1 modulates the unfolded protein response by negatively regulating ATF6 stability and by tuning IRE1 signaling [#25, #26].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established EDEM1's core role by showing it extracts terminally misfolded glycoproteins from the calnexin cycle, answering how doomed clients are diverted from futile folding into degradation.\",\n      \"evidence\": \"Overexpression/knockdown with pulse-chase of misfolded glycoprotein fate and reciprocal co-IP mapping calnexin binding to the transmembrane region\",\n      \"pmids\": [\"12610306\", \"12610305\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not establish the biochemical signal distinguishing misfolded from folding-competent clients\", \"Catalytic activity not yet demonstrated\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Resolved that EDEM1 also exists as a soluble ER-lumenal species and defined a stress-regulated paralog EDEM2, clarifying that EDEM function is not restricted to a membrane-anchored form.\",\n      \"evidence\": \"Subcellular fractionation, pulse-chase ERAD assays, and XBP1 reporter assays\",\n      \"pmids\": [\"15579471\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Relationship between soluble and membrane forms unresolved\", \"Functional distinction between EDEM1 and EDEM2 substrates unaddressed\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Dissected two separable EDEM1 activities by catalytic-site mutagenesis, showing de-mannosylation and anti-aggregation chaperone function are mechanistically distinct.\",\n      \"evidence\": \"Site-directed mutagenesis of the conserved alpha-1,2-mannosidase catalytic residue (E220Q) with N-glycan analysis and non-reducing SDS-PAGE; comparison against control ER proteins\",\n      \"pmids\": [\"16987498\", \"16629899\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct enzymatic activity of purified protein not shown\", \"Molecular basis of anti-aggregation function undefined\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Revealed a non-canonical ER exit route by localizing endogenous EDEM1 to non-COPII ~150 nm vesicles (EDEMosomes) shared with Derlin-2 and misfolded substrate.\",\n      \"evidence\": \"Quantitative immunogold electron microscopy with serial sections and subcellular fractionation\",\n      \"pmids\": [\"17360537\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Machinery generating these vesicles unidentified\", \"Destination and functional purpose of vesicular EDEM1 unresolved\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Decoupled substrate recognition from glycan chemistry, showing EDEM1 binds nonnative proteins glycan-independently while its mannosidase-like domain targets clients to SEL1L.\",\n      \"evidence\": \"Co-IP under kifunensine, mannosidase-domain mutants, and SEL1L pulldowns; extended by an opsin chaperone study and the first demonstration of basal autophagic turnover of EDEM1\",\n      \"pmids\": [\"19524542\", \"19934218\", \"19266160\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The protein determinant recognized by EDEM1 not yet defined\", \"How recognition and SEL1L handoff are coordinated unclear\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Placed EDEM1 upstream of mannose-dependent lectins and showed its overexpression can bypass the ER mannosidase I requirement, refining the ERAD step order.\",\n      \"evidence\": \"ERManI knockdown with EDEM1 overexpression, deletion mutants lacking the carbohydrate-recognition domain, and colocalization at the pericentriolar ERQC; isoform analysis defining soluble-ERdj5 vs membrane-SEL1L partitioning\",\n      \"pmids\": [\"21062743\", \"21917589\", \"21632540\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Quantitative contribution of mannose-dependent vs -independent routing in vivo unresolved\", \"Regulation of isoform ratio not defined\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Generalized EDEM1's client repertoire to nonglycosylated proteins and toxin chains, and identified hydrophobicity and a disordered N-terminal region as the recognition basis.\",\n      \"evidence\": \"Domain deletion and substrate-hydrophobicity mutagenesis with co-IP and ERAD/retrotranslocation assays across multiple substrates (NHK, H2a, ricin A-chain, BACE457, tyrosinase)\",\n      \"pmids\": [\"23233672\", \"22905195\", \"25655076\", \"21388347\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural details of disordered-region engagement unresolved\", \"Whether the same surface binds glycosylated and nonglycosylated clients unclear\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstrated intrinsic mannosidase activity directly in vitro and showed it is substrate-state dependent and enhanced by oxidoreductase partners, and that client capture involves redox-sensitive thiol contacts.\",\n      \"evidence\": \"In vitro mannosidase assays with purified EDEM1/2 on native vs denatured substrates, co-IP with TXNDC11; reducing/non-reducing co-IP with client Cys256 mutants and IDR/MLD constructs\",\n      \"pmids\": [\"30374462\", \"30021839\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and dynamics of oxidoreductase coupling in cells unresolved\", \"How redox state gates substrate release undefined\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined the precise enzymatic step EDEM1 catalyzes (M8B to M7/M6/M5) using reconstituted purified enzyme, situating it alongside EDEM3 in N-glycan trimming.\",\n      \"evidence\": \"In vitro mannosidase assay with purified EDEM1 and EDEM3 on pyridylamine-labeled M8B and glycoprotein substrates with HPLC profiling\",\n      \"pmids\": [\"34698634\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional division of labor between EDEM1 and EDEM3 in vivo unresolved\", \"Kinetic parameters on physiological clients undefined\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended EDEM1 to physiological and disease-relevant clients (APP, EGFR, TSP1) and to UPR regulation, showing roles beyond bulk misfolded-protein clearance.\",\n      \"evidence\": \"Knockdown/overexpression with co-IP, pulse-chase, Abeta ELISA, and maturation/localization readouts; ATF6 export/cleavage assays and IRE1/JNK/c-Jun pathway analysis with insulin secretion in beta cells and islets\",\n      \"pmids\": [\"35008544\", \"37569550\", \"30281916\", \"37822496\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect effects on UPR transducers not fully separated\", \"Physiological significance of individual substrate effects in tissue not established\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Mapped how EDEM1 itself is degraded, showing dual ERAD (SEL1L/HRD1, YOD1, XTP3-B, ERdj3, VIMP, BAG6, JB12) and selective autophagy routes, establishing autoregulation of the ERAD machinery.\",\n      \"evidence\": \"Sibling knockdowns of ERAD components, proteasome and autophagy inhibitors, pulse-chase; PNGase-dependent p62/NBR1 routing imaging and autoregulatory complex mass spectrometry\",\n      \"pmids\": [\"38682256\", \"24664425\", \"32423001\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Trigger selecting ERAD vs autophagy for EDEM1 turnover unresolved\", \"Physiological consequences of dysregulated EDEM1 levels undefined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Provided the first structural model of EDEM client engagement, showing the EDEM catalytic domain nesting in a PDI arc with disulfide-mediated client capture and a redox relay priming PDI as the ERAD reductase.\",\n      \"evidence\": \"Cryo-EM of EDEM:PDI heterodimer with/without A1AT-NHK client (thermophilic orthologs), mass spectrometry of disulfide bonds, and in vitro redox assays (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.01.29.635535\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Awaits peer review and confirmation in human EDEM1\", \"How redox cycling couples to retrotranslocation in the full dislocon unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how EDEM1 integrates substrate recognition, catalytic trimming, redox chemistry, and isoform/vesicle partitioning into a single regulated decision to commit a client to degradation in vivo.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No quantitative model linking mannose-dependent and -independent routing\", \"Physiological regulation of EDEMosome formation and EDEM1 turnover unknown\", \"No high-resolution human EDEM1 structure with client\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140098\", \"supporting_discovery_ids\": [9, 18, 21]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [9, 18, 21]},\n      {\"term_id\": \"GO:0044183\", \"supporting_discovery_ids\": [3, 4]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [6, 14, 19]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [2, 5, 12]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 1, 6, 24]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [25, 26]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [8, 16, 20]}\n    ],\n    \"complexes\": [\"SEL1L/HRD1 ERAD dislocation complex\", \"EDEM:PDI heterodimer\"],\n    \"partners\": [\"CANX\", \"SEL1L\", \"DNAJC10\", \"TXNDC11\", \"SQSTM1\", \"NBR1\", \"PDIA\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}