{"gene":"STIMATE","run_date":"2026-06-10T07:46:43","timeline":{"discoveries":[{"year":2015,"finding":"STIMATE (encoded by TMEM110) is an ER-resident multi-transmembrane protein that physically interacts with STIM1 to promote STIM1 conformational switch; genetic depletion of STIMATE substantially reduces STIM1 puncta formation at ER-PM junctions and suppresses Ca²⁺-NFAT signalling, identifying it as a positive regulator of store-operated Ca²⁺ entry.","method":"Spatially restricted biotin labelling in situ coupled with mass spectrometry, bimolecular fluorescence complementation, Co-IP/pulldown, live-cell imaging, genetic knockdown with Ca²⁺ flux and NFAT reporter assays","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (proteomics, BiFC, Co-IP, live imaging, functional Ca²⁺/NFAT assays) in a single rigorous study, confirmed by independent replication (PMID:26644574)","pmids":["26322679"],"is_preprint":false},{"year":2015,"finding":"TMEM110 (STIMATE) regulates both the long-term maintenance of ER-plasma membrane junctions and their short-term physiological remodeling during store-dependent calcium signaling, as demonstrated by RNAi-mediated depletion affecting STIM-ORAI-competent junction architecture.","method":"RNAi knockdown, live-cell imaging of ER-PM junction dynamics, Ca²⁺ flux assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — independent replication of STIMATE/TMEM110 function at ER-PM junctions using genetic depletion and functional imaging, corroborating PMID:26322679","pmids":["26644574"],"is_preprint":false},{"year":2022,"finding":"The polybasic C-terminal tail of STIMATE (STIMATE-CT) is intrinsically disordered and specifically interacts with PI(4,5)P₂- or PI(3,4,5)P₃-containing membranes; NMR titration and mutagenesis identified residues 242VRYR245 and 284KKKK287 as essential for membrane binding.","method":"Liposome pulldown assay, NMR spectroscopy, NMR titration with bicelles, site-directed mutagenesis","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution (liposome pulldown), NMR structural analysis, and mutagenesis all in one study; single lab but three orthogonal methods","pmids":["35121178"],"is_preprint":false},{"year":2026,"finding":"TMEM110 controls the conformational transition of the lysosomal ion channel TRPML1 to generate localized calcium efflux sites at lysosomes, preventing calcium overload, lysosomal membrane disruption, and leakage of mitochondrial DNA into the cytosol; loss of TMEM110 impairs mitochondrial clearance and exacerbates autoimmune tissue injury. Additionally, STING restrains TMEM110 expression under basal conditions and a naturally occurring TMEM110-STING interface mutation causes defective lysosomal DNA disposal and aberrant type I interferon activity.","method":"Conditional/genetic knockout mice, lysosomal Ca²⁺ imaging, conformational assays of TRPML1, mtDNA leakage detection, STING interaction/interface mutation analysis, traumatic brain injury model","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO with defined cellular and in vivo phenotype, functional channel conformation assay, mutation analysis, and pathway placement in a single rigorous study","pmids":["41540067"],"is_preprint":false},{"year":2023,"finding":"Exosomal STIMATE derived from type II alveolar epithelial cells is taken up by tissue-resident alveolar macrophages (TRAMs) to regulate high Ca²⁺ responsiveness and long-term Ca²⁺ signal transduction, maintaining M2-like immunophenotype and metabolic selection via the calcineurin (CaN)-PGC-1α pathway mediating mitochondrial biogenesis; conditional knockout of STIMATE in AEC-IIs leads to imbalanced immune/metabolic status of TRAMs, spontaneous inflammatory injury, and respiratory disorders.","method":"Conditional knockout mice (AEC-II-specific), exosome isolation and uptake assays, Ca²⁺ signalling measurements, metabolic profiling, calcineurin-PGC-1α pathway analysis, bleomycin fibrosis model","journal":"Theranostics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — conditional KO with defined cellular phenotype and pathway placement (CaN-PGC-1α), multiple readouts, but single lab","pmids":["36793853"],"is_preprint":false},{"year":2025,"finding":"Melatonin-induced upregulation of TMEM110 (STIMATE) participates in autophagy initiation and enhancement of osteogenic differentiation in inflamed periodontal ligament stem cells (Inf-PDLSCs).","method":"In vitro PDLSCs osteogenesis assays, autophagy assays, TMEM110 knockdown/overexpression, in vivo periodontitis model","journal":"Journal of pineal research","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — KD/OE with phenotypic readout (autophagy and osteogenesis), single lab, limited mechanistic detail in abstract","pmids":["40065592"],"is_preprint":false},{"year":2026,"finding":"Mustn1 binding to STIMATE impairs STIMATE's role in store-operated Ca²⁺ entry (SOCE); overexpression of the STIMATE-Mustn1 fusion protein reduces STIMATE's enhancement of SOCE and alters co-localization between Orai1 and STIM1. STIMATE overexpression arrests the cell cycle of MEG-01 cells in G2 phase, but the STIMATE-Mustn1 fusion does not alter the cell cycle.","method":"Overexpression of STIMATE and STIMATE-Mustn1 fusion in MEG-01 and HEK293 cells, Ca²⁺ flux (SOCE) assays, confocal imaging of Orai1/STIM1 co-localization, cell cycle analysis","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — functional Ca²⁺ assays and imaging with OE constructs in two cell lines, single lab, mechanistic detail moderate","pmids":["41677464"],"is_preprint":false}],"current_model":"STIMATE (TMEM110) is an ER-resident multi-transmembrane protein that localizes to ER-plasma membrane junctions, where it physically interacts with STIM1 (via its polybasic, intrinsically disordered C-terminal tail that also binds PI(4,5)P₂/PI(3,4,5)P₃) to promote STIM1 conformational switching and puncta formation, thereby positively regulating store-operated Ca²⁺ entry and downstream Ca²⁺-NFAT signalling; it also maintains ER-PM junction architecture, controls lysosomal function by modulating TRPML1 conformation to prevent mtDNA leakage and aberrant interferon signalling (with STING acting as a negative regulator of TMEM110 expression), and in epithelial-derived exosomes regulates macrophage Ca²⁺ signalling and metabolic reprogramming via the calcineurin-PGC-1α pathway."},"narrative":{"mechanistic_narrative":"STIMATE (encoded by TMEM110) is an ER-resident multi-transmembrane protein that acts as a positive regulator of store-operated Ca²⁺ entry by physically interacting with STIM1 to promote its conformational switch and puncta formation at ER-plasma membrane junctions, driving downstream Ca²⁺-NFAT signalling [PMID:26322679]. Beyond regulating individual STIM1 molecules, STIMATE maintains the long-term architecture of STIM-ORAI-competent ER-PM junctions and governs their physiological remodeling during store-dependent Ca²⁺ signalling [PMID:26644574]. Its membrane engagement is mediated by a polybasic, intrinsically disordered C-terminal tail that binds PI(4,5)P₂- and PI(3,4,5)P₃-containing membranes through the residues 242VRYR245 and 284KKKK287 [PMID:35121178]. STIMATE function in SOCE is antagonized by Mustn1 binding, which reduces its enhancement of SOCE and alters Orai1/STIM1 co-localization [PMID:41677464]. STIMATE also operates at the lysosome, where it controls the conformational transition of the ion channel TRPML1 to generate localized Ca²⁺ efflux sites, preventing lysosomal Ca²⁺ overload, membrane disruption, and cytosolic leakage of mitochondrial DNA; its loss impairs mitochondrial clearance and exacerbates autoimmune tissue injury, with STING restraining TMEM110 expression and a TMEM110-STING interface mutation causing aberrant type I interferon activity [PMID:41540067]. In addition, exosomal STIMATE secreted by alveolar epithelial cells is taken up by tissue-resident alveolar macrophages to sustain Ca²⁺ responsiveness and M2-like immunometabolic programming via the calcineurin-PGC-1α pathway [PMID:36793853].","teleology":[{"year":2015,"claim":"Established STIMATE as a dedicated positive regulator of store-operated Ca²⁺ entry, answering how STIM1 activation is potentiated at ER-PM junctions.","evidence":"Spatially restricted biotin proximity labelling/MS, BiFC, Co-IP, live-cell imaging, and Ca²⁺/NFAT reporter assays with genetic depletion","pmids":["26322679"],"confidence":"High","gaps":["Structural basis of the STIMATE-STIM1 interaction not resolved","Did not define how STIMATE itself is regulated"]},{"year":2015,"claim":"Showed STIMATE controls both maintenance and dynamic remodeling of ER-PM junction architecture, distinguishing a structural role from direct STIM1 binding.","evidence":"RNAi depletion with live-cell imaging of ER-PM junction dynamics and Ca²⁺ flux assays","pmids":["26644574"],"confidence":"High","gaps":["Molecular determinants of junction tethering not mapped","Relationship to other junction-resident proteins unaddressed"]},{"year":2022,"claim":"Defined the membrane-binding mechanism of STIMATE, showing its disordered polybasic C-terminal tail engages specific phosphoinositides via identified residues.","evidence":"Liposome pulldown, NMR spectroscopy/titration with bicelles, and site-directed mutagenesis in vitro","pmids":["35121178"],"confidence":"High","gaps":["Functional consequence of phosphoinositide binding in cells not tested","Single-lab in vitro reconstitution"]},{"year":2023,"claim":"Revealed a non-cell-autonomous, exosome-mediated role for STIMATE in shaping macrophage Ca²⁺ signalling and immunometabolism between epithelial and immune cells.","evidence":"AEC-II-specific conditional knockout mice, exosome isolation/uptake assays, Ca²⁺ and metabolic profiling, calcineurin-PGC-1α pathway analysis, bleomycin model","pmids":["36793853"],"confidence":"Medium","gaps":["Mechanism of STIMATE loading into exosomes unknown","Single lab; how exosomal STIMATE acts in recipient cells not molecularly resolved"]},{"year":2025,"claim":"Linked STIMATE to autophagy and osteogenic differentiation under inflammatory conditions, broadening its functional repertoire beyond Ca²⁺ junction biology.","evidence":"In vitro PDLSC osteogenesis and autophagy assays with TMEM110 knockdown/overexpression and an in vivo periodontitis model","pmids":["40065592"],"confidence":"Medium","gaps":["Mechanistic link between STIMATE and autophagy machinery not defined","Limited mechanistic detail available"]},{"year":2026,"claim":"Extended STIMATE function to the lysosome, showing it regulates TRPML1 conformation to prevent mtDNA leakage and aberrant interferon signalling, with STING as an upstream restraint.","evidence":"Conditional/genetic knockout mice, lysosomal Ca²⁺ imaging, TRPML1 conformational assays, mtDNA leakage detection, STING interface mutation analysis, TBI model","pmids":["41540067"],"confidence":"High","gaps":["Direct physical mechanism of TRPML1 conformational control not resolved","How STING transcriptionally/post-transcriptionally restrains TMEM110 unclear"]},{"year":2026,"claim":"Identified Mustn1 as a negative regulator of STIMATE-driven SOCE and a modulator of its effect on cell cycle progression.","evidence":"Overexpression of STIMATE and STIMATE-Mustn1 fusion in MEG-01/HEK293 cells, SOCE assays, Orai1/STIM1 co-localization imaging, cell cycle analysis","pmids":["41677464"],"confidence":"Medium","gaps":["Endogenous Mustn1-STIMATE interaction relied on fusion overexpression","Mechanism by which STIMATE arrests cell cycle in G2 not defined"]},{"year":null,"claim":"How STIMATE's distinct activities at ER-PM junctions, lysosomes, and in exosomal signalling are coordinated and regulated within a single cell remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model integrating STIM1 and TRPML1 engagement","Regulatory logic linking phosphoinositide binding to each functional context unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,6]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[2]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[0,1]},{"term_id":"GO:0005764","term_label":"lysosome","supporting_discovery_ids":[3]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[3,4]}],"complexes":[],"partners":["STIM1","TRPML1","STING","MUSTN1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q86TL2","full_name":"Store-operated calcium entry regulator STIMATE","aliases":["STIM-activating enhancer encoded by TMEM110","Transmembrane protein 110"],"length_aa":294,"mass_kda":33.2,"function":"Acts as a regulator of store-operated Ca(2+) entry (SOCE) at junctional sites that connect the endoplasmic reticulum (ER) and plasma membrane (PM), called ER-plasma membrane (ER-PM) junction or cortical ER (PubMed:26322679, PubMed:26644574). SOCE is a Ca(2+) influx following depletion of intracellular Ca(2+) stores (PubMed:26322679). Acts by interacting with STIM1, promoting STIM1 conformational switch (PubMed:26322679). Involved in STIM1 relocalization to ER-PM junctions (PubMed:26644574). Contributes to the maintenance and reorganization of store-dependent ER-PM junctions (PubMed:26644574)","subcellular_location":"Endoplasmic reticulum membrane","url":"https://www.uniprot.org/uniprotkb/Q86TL2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/STIMATE","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/STIMATE","total_profiled":1310},"omim":[{"mim_id":"617189","title":"TRANSMEMBRANE PROTEIN 110; TMEM110","url":"https://www.omim.org/entry/617189"},{"mim_id":"605921","title":"STROMAL INTERACTION MOLECULE 1; STIM1","url":"https://www.omim.org/entry/605921"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/STIMATE"},"hgnc":{"alias_symbol":["MGC52022"],"prev_symbol":["TMEM110"]},"alphafold":{"accession":"Q86TL2","domains":[{"cath_id":"-","chopping":"35-63_75-119_163-221","consensus_level":"high","plddt":84.8343,"start":35,"end":221}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86TL2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q86TL2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q86TL2-F1-predicted_aligned_error_v6.png","plddt_mean":68.94},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=STIMATE","jax_strain_url":"https://www.jax.org/strain/search?query=STIMATE"},"sequence":{"accession":"Q86TL2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q86TL2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q86TL2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86TL2"}},"corpus_meta":[{"pmid":"26322679","id":"PMC_26322679","title":"Proteomic mapping of ER-PM junctions identifies STIMATE as a regulator of Ca²⁺ influx.","date":"2015","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/26322679","citation_count":183,"is_preprint":false},{"pmid":"26644574","id":"PMC_26644574","title":"TMEM110 regulates the maintenance and remodeling of mammalian ER-plasma membrane junctions competent for STIM-ORAI signaling.","date":"2015","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/26644574","citation_count":66,"is_preprint":false},{"pmid":"27130253","id":"PMC_27130253","title":"Molecular modulators of store-operated calcium entry.","date":"2016","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/27130253","citation_count":60,"is_preprint":false},{"pmid":"38631091","id":"PMC_38631091","title":"Impact of ambient air pollution on colorectal cancer risk and survival: insights from a prospective cohort and epigenetic Mendelian randomization study.","date":"2024","source":"EBioMedicine","url":"https://pubmed.ncbi.nlm.nih.gov/38631091","citation_count":50,"is_preprint":false},{"pmid":"28731711","id":"PMC_28731711","title":"Identification of Proteomic Features To Distinguish Benign Pulmonary Nodules from Lung Adenocarcinoma.","date":"2017","source":"Journal of proteome research","url":"https://pubmed.ncbi.nlm.nih.gov/28731711","citation_count":45,"is_preprint":false},{"pmid":"36793853","id":"PMC_36793853","title":"Exosomal STIMATE derived from type II alveolar epithelial cells controls metabolic reprogramming of tissue-resident alveolar macrophages.","date":"2023","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/36793853","citation_count":38,"is_preprint":false},{"pmid":"27399837","id":"PMC_27399837","title":"Loss of all 3 Extended Synaptotagmins does not affect normal mouse development, viability or fertility.","date":"2016","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/27399837","citation_count":35,"is_preprint":false},{"pmid":"25465359","id":"PMC_25465359","title":"Genome-wide association study for semen volume and total number of sperm in Holstein-Friesian bulls.","date":"2014","source":"Animal reproduction science","url":"https://pubmed.ncbi.nlm.nih.gov/25465359","citation_count":35,"is_preprint":false},{"pmid":"36203605","id":"PMC_36203605","title":"Immunological risk factors for sepsis-associated delirium and mortality in ICU patients.","date":"2022","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/36203605","citation_count":25,"is_preprint":false},{"pmid":"18312368","id":"PMC_18312368","title":"Biologic response to subcutaneous and intranasal therapy with desmopressin in a large Amish kindred with Type 2M von Willebrand disease.","date":"2008","source":"Haemophilia : the official journal of the World Federation of Hemophilia","url":"https://pubmed.ncbi.nlm.nih.gov/18312368","citation_count":8,"is_preprint":false},{"pmid":"40065592","id":"PMC_40065592","title":"Melatonin Increased Autophagy Level to Facilitate Osteogenesis of Inflamed PDLSCs Through TMEM110 Signaling Pathways.","date":"2025","source":"Journal of pineal research","url":"https://pubmed.ncbi.nlm.nih.gov/40065592","citation_count":4,"is_preprint":false},{"pmid":"28974198","id":"PMC_28974198","title":"Integrated pipeline for inferring the evolutionary history of a gene family embedded in the species tree: a case study on the STIMATE gene family.","date":"2017","source":"BMC bioinformatics","url":"https://pubmed.ncbi.nlm.nih.gov/28974198","citation_count":4,"is_preprint":false},{"pmid":"39278114","id":"PMC_39278114","title":"Single-nucleus RNA sequencing reveals cell types, genes, and regulatory factors influencing melanogenesis in the breast muscle of Xuefeng black-bone chicken.","date":"2024","source":"Poultry science","url":"https://pubmed.ncbi.nlm.nih.gov/39278114","citation_count":3,"is_preprint":false},{"pmid":"35121178","id":"PMC_35121178","title":"Biochemical and NMR studies reveal specific interaction between STIMATE C-tail and PI(4,5)P2 or PI(3,4,5)P3-containing membrane.","date":"2022","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/35121178","citation_count":2,"is_preprint":false},{"pmid":"37859957","id":"PMC_37859957","title":"Cross-species analysis of differential transcript usage in humans and chickens with fatty liver disease.","date":"2023","source":"Veterinary world","url":"https://pubmed.ncbi.nlm.nih.gov/37859957","citation_count":2,"is_preprint":false},{"pmid":"41540067","id":"PMC_41540067","title":"Tmem110 regulates the conformation of TRPML1 to maintain endolysosomal homeostasis and prevent mitochondrial DNA leakage and pathological self-DNA processing.","date":"2026","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/41540067","citation_count":1,"is_preprint":false},{"pmid":"40242870","id":"PMC_40242870","title":"Epigenetic Modulation of Vascular Smooth Muscle Cell Phenotype Switching in Early-Onset Acute Myocardial Infarction.","date":"2025","source":"Arteriosclerosis, thrombosis, and vascular biology","url":"https://pubmed.ncbi.nlm.nih.gov/40242870","citation_count":1,"is_preprint":false},{"pmid":"41677464","id":"PMC_41677464","title":"STIMATE-Mustn1 Fusion Protein Prevents Ca2+ Overload in Cells That Present Increased SOCE.","date":"2026","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/41677464","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11445,"output_tokens":2193,"usd":0.033615,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9264,"output_tokens":2780,"usd":0.05791,"stage2_stop_reason":"end_turn"},"total_usd":0.091525,"stage1_batch_id":"msgbatch_013LWZZT6MWaqB9qyTSeQBmU","stage2_batch_id":"msgbatch_0194ZzQaTGNpruoNJBbEMbt3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2015,\n      \"finding\": \"STIMATE (encoded by TMEM110) is an ER-resident multi-transmembrane protein that physically interacts with STIM1 to promote STIM1 conformational switch; genetic depletion of STIMATE substantially reduces STIM1 puncta formation at ER-PM junctions and suppresses Ca²⁺-NFAT signalling, identifying it as a positive regulator of store-operated Ca²⁺ entry.\",\n      \"method\": \"Spatially restricted biotin labelling in situ coupled with mass spectrometry, bimolecular fluorescence complementation, Co-IP/pulldown, live-cell imaging, genetic knockdown with Ca²⁺ flux and NFAT reporter assays\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (proteomics, BiFC, Co-IP, live imaging, functional Ca²⁺/NFAT assays) in a single rigorous study, confirmed by independent replication (PMID:26644574)\",\n      \"pmids\": [\"26322679\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"TMEM110 (STIMATE) regulates both the long-term maintenance of ER-plasma membrane junctions and their short-term physiological remodeling during store-dependent calcium signaling, as demonstrated by RNAi-mediated depletion affecting STIM-ORAI-competent junction architecture.\",\n      \"method\": \"RNAi knockdown, live-cell imaging of ER-PM junction dynamics, Ca²⁺ flux assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — independent replication of STIMATE/TMEM110 function at ER-PM junctions using genetic depletion and functional imaging, corroborating PMID:26322679\",\n      \"pmids\": [\"26644574\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The polybasic C-terminal tail of STIMATE (STIMATE-CT) is intrinsically disordered and specifically interacts with PI(4,5)P₂- or PI(3,4,5)P₃-containing membranes; NMR titration and mutagenesis identified residues 242VRYR245 and 284KKKK287 as essential for membrane binding.\",\n      \"method\": \"Liposome pulldown assay, NMR spectroscopy, NMR titration with bicelles, site-directed mutagenesis\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution (liposome pulldown), NMR structural analysis, and mutagenesis all in one study; single lab but three orthogonal methods\",\n      \"pmids\": [\"35121178\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"TMEM110 controls the conformational transition of the lysosomal ion channel TRPML1 to generate localized calcium efflux sites at lysosomes, preventing calcium overload, lysosomal membrane disruption, and leakage of mitochondrial DNA into the cytosol; loss of TMEM110 impairs mitochondrial clearance and exacerbates autoimmune tissue injury. Additionally, STING restrains TMEM110 expression under basal conditions and a naturally occurring TMEM110-STING interface mutation causes defective lysosomal DNA disposal and aberrant type I interferon activity.\",\n      \"method\": \"Conditional/genetic knockout mice, lysosomal Ca²⁺ imaging, conformational assays of TRPML1, mtDNA leakage detection, STING interaction/interface mutation analysis, traumatic brain injury model\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with defined cellular and in vivo phenotype, functional channel conformation assay, mutation analysis, and pathway placement in a single rigorous study\",\n      \"pmids\": [\"41540067\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Exosomal STIMATE derived from type II alveolar epithelial cells is taken up by tissue-resident alveolar macrophages (TRAMs) to regulate high Ca²⁺ responsiveness and long-term Ca²⁺ signal transduction, maintaining M2-like immunophenotype and metabolic selection via the calcineurin (CaN)-PGC-1α pathway mediating mitochondrial biogenesis; conditional knockout of STIMATE in AEC-IIs leads to imbalanced immune/metabolic status of TRAMs, spontaneous inflammatory injury, and respiratory disorders.\",\n      \"method\": \"Conditional knockout mice (AEC-II-specific), exosome isolation and uptake assays, Ca²⁺ signalling measurements, metabolic profiling, calcineurin-PGC-1α pathway analysis, bleomycin fibrosis model\",\n      \"journal\": \"Theranostics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional KO with defined cellular phenotype and pathway placement (CaN-PGC-1α), multiple readouts, but single lab\",\n      \"pmids\": [\"36793853\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Melatonin-induced upregulation of TMEM110 (STIMATE) participates in autophagy initiation and enhancement of osteogenic differentiation in inflamed periodontal ligament stem cells (Inf-PDLSCs).\",\n      \"method\": \"In vitro PDLSCs osteogenesis assays, autophagy assays, TMEM110 knockdown/overexpression, in vivo periodontitis model\",\n      \"journal\": \"Journal of pineal research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — KD/OE with phenotypic readout (autophagy and osteogenesis), single lab, limited mechanistic detail in abstract\",\n      \"pmids\": [\"40065592\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Mustn1 binding to STIMATE impairs STIMATE's role in store-operated Ca²⁺ entry (SOCE); overexpression of the STIMATE-Mustn1 fusion protein reduces STIMATE's enhancement of SOCE and alters co-localization between Orai1 and STIM1. STIMATE overexpression arrests the cell cycle of MEG-01 cells in G2 phase, but the STIMATE-Mustn1 fusion does not alter the cell cycle.\",\n      \"method\": \"Overexpression of STIMATE and STIMATE-Mustn1 fusion in MEG-01 and HEK293 cells, Ca²⁺ flux (SOCE) assays, confocal imaging of Orai1/STIM1 co-localization, cell cycle analysis\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — functional Ca²⁺ assays and imaging with OE constructs in two cell lines, single lab, mechanistic detail moderate\",\n      \"pmids\": [\"41677464\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"STIMATE (TMEM110) is an ER-resident multi-transmembrane protein that localizes to ER-plasma membrane junctions, where it physically interacts with STIM1 (via its polybasic, intrinsically disordered C-terminal tail that also binds PI(4,5)P₂/PI(3,4,5)P₃) to promote STIM1 conformational switching and puncta formation, thereby positively regulating store-operated Ca²⁺ entry and downstream Ca²⁺-NFAT signalling; it also maintains ER-PM junction architecture, controls lysosomal function by modulating TRPML1 conformation to prevent mtDNA leakage and aberrant interferon signalling (with STING acting as a negative regulator of TMEM110 expression), and in epithelial-derived exosomes regulates macrophage Ca²⁺ signalling and metabolic reprogramming via the calcineurin-PGC-1α pathway.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"STIMATE (encoded by TMEM110) is an ER-resident multi-transmembrane protein that acts as a positive regulator of store-operated Ca²⁺ entry by physically interacting with STIM1 to promote its conformational switch and puncta formation at ER-plasma membrane junctions, driving downstream Ca²⁺-NFAT signalling [#0]. Beyond regulating individual STIM1 molecules, STIMATE maintains the long-term architecture of STIM-ORAI-competent ER-PM junctions and governs their physiological remodeling during store-dependent Ca²⁺ signalling [#1]. Its membrane engagement is mediated by a polybasic, intrinsically disordered C-terminal tail that binds PI(4,5)P₂- and PI(3,4,5)P₃-containing membranes through the residues 242VRYR245 and 284KKKK287 [#2]. STIMATE function in SOCE is antagonized by Mustn1 binding, which reduces its enhancement of SOCE and alters Orai1/STIM1 co-localization [#6]. STIMATE also operates at the lysosome, where it controls the conformational transition of the ion channel TRPML1 to generate localized Ca²⁺ efflux sites, preventing lysosomal Ca²⁺ overload, membrane disruption, and cytosolic leakage of mitochondrial DNA; its loss impairs mitochondrial clearance and exacerbates autoimmune tissue injury, with STING restraining TMEM110 expression and a TMEM110-STING interface mutation causing aberrant type I interferon activity [#3]. In addition, exosomal STIMATE secreted by alveolar epithelial cells is taken up by tissue-resident alveolar macrophages to sustain Ca²⁺ responsiveness and M2-like immunometabolic programming via the calcineurin-PGC-1α pathway [#4].\",\n  \"teleology\": [\n    {\n      \"year\": 2015,\n      \"claim\": \"Established STIMATE as a dedicated positive regulator of store-operated Ca²⁺ entry, answering how STIM1 activation is potentiated at ER-PM junctions.\",\n      \"evidence\": \"Spatially restricted biotin proximity labelling/MS, BiFC, Co-IP, live-cell imaging, and Ca²⁺/NFAT reporter assays with genetic depletion\",\n      \"pmids\": [\"26322679\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the STIMATE-STIM1 interaction not resolved\", \"Did not define how STIMATE itself is regulated\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Showed STIMATE controls both maintenance and dynamic remodeling of ER-PM junction architecture, distinguishing a structural role from direct STIM1 binding.\",\n      \"evidence\": \"RNAi depletion with live-cell imaging of ER-PM junction dynamics and Ca²⁺ flux assays\",\n      \"pmids\": [\"26644574\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular determinants of junction tethering not mapped\", \"Relationship to other junction-resident proteins unaddressed\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined the membrane-binding mechanism of STIMATE, showing its disordered polybasic C-terminal tail engages specific phosphoinositides via identified residues.\",\n      \"evidence\": \"Liposome pulldown, NMR spectroscopy/titration with bicelles, and site-directed mutagenesis in vitro\",\n      \"pmids\": [\"35121178\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of phosphoinositide binding in cells not tested\", \"Single-lab in vitro reconstitution\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Revealed a non-cell-autonomous, exosome-mediated role for STIMATE in shaping macrophage Ca²⁺ signalling and immunometabolism between epithelial and immune cells.\",\n      \"evidence\": \"AEC-II-specific conditional knockout mice, exosome isolation/uptake assays, Ca²⁺ and metabolic profiling, calcineurin-PGC-1α pathway analysis, bleomycin model\",\n      \"pmids\": [\"36793853\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of STIMATE loading into exosomes unknown\", \"Single lab; how exosomal STIMATE acts in recipient cells not molecularly resolved\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Linked STIMATE to autophagy and osteogenic differentiation under inflammatory conditions, broadening its functional repertoire beyond Ca²⁺ junction biology.\",\n      \"evidence\": \"In vitro PDLSC osteogenesis and autophagy assays with TMEM110 knockdown/overexpression and an in vivo periodontitis model\",\n      \"pmids\": [\"40065592\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanistic link between STIMATE and autophagy machinery not defined\", \"Limited mechanistic detail available\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Extended STIMATE function to the lysosome, showing it regulates TRPML1 conformation to prevent mtDNA leakage and aberrant interferon signalling, with STING as an upstream restraint.\",\n      \"evidence\": \"Conditional/genetic knockout mice, lysosomal Ca²⁺ imaging, TRPML1 conformational assays, mtDNA leakage detection, STING interface mutation analysis, TBI model\",\n      \"pmids\": [\"41540067\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct physical mechanism of TRPML1 conformational control not resolved\", \"How STING transcriptionally/post-transcriptionally restrains TMEM110 unclear\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Identified Mustn1 as a negative regulator of STIMATE-driven SOCE and a modulator of its effect on cell cycle progression.\",\n      \"evidence\": \"Overexpression of STIMATE and STIMATE-Mustn1 fusion in MEG-01/HEK293 cells, SOCE assays, Orai1/STIM1 co-localization imaging, cell cycle analysis\",\n      \"pmids\": [\"41677464\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Endogenous Mustn1-STIMATE interaction relied on fusion overexpression\", \"Mechanism by which STIMATE arrests cell cycle in G2 not defined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How STIMATE's distinct activities at ER-PM junctions, lysosomes, and in exosomal signalling are coordinated and regulated within a single cell remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model integrating STIM1 and TRPML1 engagement\", \"Regulatory logic linking phosphoinositide binding to each functional context unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 6]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [3, 4]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"STIM1\", \"TRPML1\", \"STING\", \"Mustn1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"tie","faith_supported":6,"faith_total":6,"faith_pct":100.0}}