{"gene":"MMP24","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":1999,"finding":"MT5-MMP (MMP24) activates progelatinase A (MMP2) when co-expressed in MDCK cells, and this activity is dependent on proteolytic activity since a Glu→Ala mutation in the zinc-binding motif (HE255LGH) abolishes activation.","method":"Co-expression in MDCK cells, active-site mutagenesis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — active-site mutagenesis with functional readout, replicated in same year by independent lab (PMID:10363975)","pmids":["10085137"],"is_preprint":false},{"year":1999,"finding":"MT5-MMP tends to shed from the cell surface as a soluble proteinase, distinguishing it from other MT-MMPs which remain membrane-bound.","method":"Cell expression and protein analysis (Western blot, conditioned media analysis)","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — demonstrated in cell expression system, consistent with independent replication","pmids":["10085137"],"is_preprint":false},{"year":1999,"finding":"Human MT5-MMP (MMP24) localizes to the plasma membrane when expressed in COS-7 cells, as shown by immunofluorescence and Western blot.","method":"Immunofluorescence and Western blot of transfected COS-7 cells","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — two orthogonal methods (IF + WB), single lab","pmids":["10363975"],"is_preprint":false},{"year":1999,"finding":"The catalytic domain of MT5-MMP cleaves progelatinase A to generate the Mr 62,000 active form, demonstrated using a GST-fusion catalytic domain expressed in E. coli.","method":"In vitro proteolytic assay with recombinant catalytic domain (GST fusion)","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with purified recombinant protein, replicated in same year (PMID:10085137)","pmids":["10363975"],"is_preprint":false},{"year":1999,"finding":"Purified soluble MT5-MMP activates progelatinase A in a TIMP-2-sensitive fashion, and preferentially cleaves proteoglycans among extracellular matrix components.","method":"Biochemical purification, in vitro proteolytic assay with ECM substrate panel","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with purified enzyme and defined substrates, single lab","pmids":["10622708"],"is_preprint":false},{"year":2004,"finding":"The C-terminal EWV motif of MT5-MMP serves as a retrieval signal for internalized MT5-MMP by interacting with the PDZ domain protein Mint-3, which recycles MT5-MMP to the plasma membrane. Deletion of the EWV signal impairs recycling and decreases surface activity. Mint-3 knockdown by siRNA decreases MT5-MMP activity.","method":"Yeast two-hybrid screening, deletion mutagenesis, siRNA knockdown, cell surface activity assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — yeast two-hybrid identification, mutagenesis of binding motif, siRNA KD, multiple orthogonal methods in one study","pmids":["14990567"],"is_preprint":false},{"year":2009,"finding":"MT5-MMP cleaves N-cadherin in peptidergic nociceptors (CGRP+ dorsal root ganglion neurons); Mmp24-deficient mice show enhanced basal thermal sensitivity and failure to develop thermal hyperalgesia during inflammation, correlating with altered N-cadherin-mediated neuro-immune cell interactions.","method":"Mmp24 knockout mouse, behavioral nociception assays, N-cadherin cleavage analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO with defined cellular and behavioral phenotype, mechanistic link to N-cadherin cleavage, multiple readouts","pmids":["19805319"],"is_preprint":false},{"year":2014,"finding":"MT5-MMP cleaves the N-cadherin ectodomain in adult neural stem cells (NSCs) and ependymocytes of the subependymal zone. MT5-MMP-mediated N-cadherin shedding is required for proper activation of NSCs under physiological and regenerative conditions but is dispensable for NSC generation and identity.","method":"MT5-MMP knockout mice, N-cadherin shedding assays, NSC functional assays in vivo","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO in vivo, specific substrate (N-cadherin) cleavage demonstrated, functional consequence on stem cell quiescence, rigorous controls","pmids":["24952463"],"is_preprint":false},{"year":2015,"finding":"MT5-MMP co-localizes and co-immunoprecipitates with APP in HEKswe cells and promotes amyloidogenic APP processing, increasing levels of Aβ, C99, and a soluble APP fragment of 95 kDa (sAPP95). MT5-MMP deficiency in 5xFAD mice reduces Aβ, C99, and sAPP95 levels without altering α-, β-, or γ-secretase activities.","method":"Co-immunoprecipitation, co-localization (immunofluorescence), MT5-MMP KO × 5xFAD cross, ELISA, Western blot","journal":"Cellular and molecular life sciences : CMLS","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP in cell model, in vivo KO mouse validation, multiple orthogonal methods","pmids":["26202697"],"is_preprint":false},{"year":2017,"finding":"MT5-MMP expressed in HEKswe cells localizes to early endosomes and promotes trafficking of APP and C99 into early endosomes, as well as increased Aβ40 in cell supernatants, establishing a pro-amyloidogenic role via endosomal trafficking.","method":"Immunofluorescence co-localization, subcellular fractionation, ELISA in transfected HEKswe cells","journal":"Frontiers in molecular neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — subcellular localization linked to functional APP trafficking consequence, single lab","pmids":["28119565"],"is_preprint":false},{"year":2021,"finding":"Deletion of C-terminal non-catalytic domains of MT5-MMP hampers its ability to process APP and release sAPP95. Catalytically inactive MT5-MMP variants still increase Aβ and promote APP/C99 sorting to the endolysosomal system via interactions of the C-terminal portion with C99. Deletion of the C-terminal domain causes proteasomal degradation of C99 and prevents Aβ accumulation.","method":"Domain deletion mutagenesis, catalytic-dead variants, subcellular fractionation, proteasome inhibitors, Western blot in HEKswe cells","journal":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — structure-function mutagenesis with multiple deletion variants, both proteolytic and non-proteolytic mechanisms dissected, single lab with multiple orthogonal methods","pmids":["34117802"],"is_preprint":false},{"year":2021,"finding":"FTO (m6A eraser) binds to Mmp24 mRNA in spinal cord neurons after spinal nerve ligation, reducing m6A modification and facilitating MMP24 translation. MMP24 upregulation in spinal cord neurons promotes ERK activation and nociceptive hypersensitivity.","method":"MeRIP-seq, RNA immunoprecipitation, FTO overexpression/knockdown in spinal neurons, ERK activation assays, neuropathic pain behavioral tests","journal":"Frontiers in pharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MeRIP-seq + RIP assay + FTO OE/KD, multiple methods, single lab","pmids":["33995105"],"is_preprint":false},{"year":2024,"finding":"PCSK6 cleaves and activates MT5-MMP by recognizing the RRRNKR sequence in the MT5-MMP N-terminal propeptide domain. Mutation or knockout of this cleavage motif prevents PCSK6 from interacting with and cleaving MT5-MMP, reducing amyloidogenic APP processing.","method":"Co-immunoprecipitation, site-directed mutagenesis of cleavage motif, PCSK6 knockdown in N2AAPP cells and AAV-mediated KD in APP23/PS45 mice","journal":"Experimental neurology","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — cleavage motif mutagenesis abolishes interaction, in vitro cell and in vivo AAV KD validation, multiple methods","pmids":["38216110"],"is_preprint":false},{"year":2024,"finding":"MT5-MMP localizes to the nucleus, cytosol, and cytosolic subcellular granules in CHO cells stably expressing APP751. Using subcellular-targeted MT5-MMP fusion proteins, η-secretase (sAPPη) activity was highest when MT5-MMP was directed to the endosome, Golgi, plasma membrane, or mitochondria.","method":"Subcellular-targeted fusion protein constructs, sAPPη ELISA, immunofluorescence localization in CHO-APP751 cells","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — targeted fusion proteins to specific organelles with functional readout, single lab","pmids":["38345408"],"is_preprint":false},{"year":2024,"finding":"Yeast two-hybrid screening identified N4BP2L1, TMX3, EIG121, BIN1, RUFY4, HTRA1, and TMEM199 as binding partners of the MT5-MMP C-terminal domain in a human brain cDNA library. Binding of N4BP2L1, EIG121, BIN1, or TMX3 resulted in significant increase in sAPPη production.","method":"Yeast two-hybrid screen, co-expression functional assay (sAPPη ELISA)","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — yeast two-hybrid identification with functional follow-up in cell model, single lab","pmids":["38345408"],"is_preprint":false},{"year":2025,"finding":"Specific amino acid mutations in the intracellular (IC) domain of MT5-MMP induce C99 degradation and decrease Aβ levels, while mutations in different IC clusters have divergent effects. The IC domain regulates subcellular trafficking of C99 through the endomembrane system and proximity with C99 as shown by proximity ligation assay.","method":"Domain deletion/substitution mutagenesis, high-content imaging, proximity ligation assay, co-transfection in human cell lines","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — mutagenesis with functional readouts and PLA, preprint not yet peer-reviewed","pmids":["bio_10.1101_2025.05.22.655451"],"is_preprint":true}],"current_model":"MT5-MMP (MMP24) is a brain-enriched membrane-type matrix metalloproteinase that acts as an η-secretase cleaving APP to generate pro-amyloidogenic fragments (Aβ, C99, sAPP95); it is activated by PCSK6-mediated cleavage of its RRRNKR propeptide motif, traffics to the cell surface via a C-terminal EWV motif interaction with the PDZ adaptor Mint-3, cleaves N-cadherin to regulate neural stem cell quiescence and nociceptor-mast cell interactions, degrades proteoglycans, and activates progelatinase A (MMP2) through its catalytic zinc-binding domain, with its non-catalytic C-terminal domains additionally controlling APP/C99 sorting through the endolysosomal system via interactions with binding partners including N4BP2L1, EIG121, BIN1, and TMX3."},"narrative":{"mechanistic_narrative":"MMP24 (MT5-MMP) is a brain-enriched membrane-type matrix metalloproteinase that couples extracellular and intracellular proteolytic functions to neural stem cell regulation, nociception, and amyloidogenic APP processing [PMID:19805319, PMID:24952463, PMID:26202697]. Its catalytic zinc-binding domain activates progelatinase A (MMP2) and preferentially degrades proteoglycans among extracellular matrix components, an activity abolished by mutation of the HEXGH zinc-binding motif and sensitive to TIMP-2 [PMID:10085137, PMID:10363975, PMID:10622708]. Surface presentation of the enzyme is controlled by recycling: its C-terminal EWV motif binds the PDZ adaptor Mint-3 to retrieve internalized MT5-MMP back to the plasma membrane, and loss of this signal or Mint-3 depletion reduces surface activity [PMID:14990567]. Activation requires PCSK6, which recognizes and cleaves the RRRNKR propeptide motif to generate active enzyme [PMID:38216110]. In neurons, MT5-MMP sheds the N-cadherin ectodomain to regulate adult neural stem cell activation and quiescence and to modulate nociceptor neuro-immune interactions, with knockout mice showing altered thermal sensitivity and impaired stem cell activation [PMID:19805319, PMID:24952463]. Acting as an η-secretase, MT5-MMP associates with APP and promotes amyloidogenic processing, raising Aβ, C99, and sAPP95 without altering α-, β-, or γ-secretase activities, in part by directing APP and C99 into the endolysosomal system [PMID:26202697, PMID:28119565, PMID:34117802]. This pro-amyloidogenic activity is only partly proteolytic: the C-terminal non-catalytic and intracellular domains govern C99 sorting and stability through brain-derived binding partners including N4BP2L1, EIG121, BIN1, and TMX3, and deletion of these domains routes C99 to proteasomal degradation [PMID:34117802, PMID:38345408, PMID:bio_10.1101_2025.05.22.655451].","teleology":[{"year":1999,"claim":"Established MT5-MMP as a catalytically active metalloproteinase capable of activating a downstream proteinase, defining its core enzymatic function and zinc-dependent mechanism.","evidence":"Co-expression in MDCK cells with active-site (Glu→Ala) mutagenesis, plus in vitro assays with recombinant catalytic domain and purified soluble enzyme","pmids":["10085137","10363975","10622708"],"confidence":"High","gaps":["Physiological MMP2 activation in vivo not demonstrated","Full ECM substrate repertoire beyond proteoglycans not defined"]},{"year":1999,"claim":"Localized MT5-MMP to the plasma membrane and showed it can shed as a soluble proteinase, distinguishing it from membrane-retained MT-MMPs.","evidence":"Immunofluorescence and Western blot of transfected COS-7 and conditioned-media analysis","pmids":["10363975","10085137"],"confidence":"Medium","gaps":["Shedding mechanism and protease responsible not identified","Relative balance of membrane vs soluble pools in neurons unknown"]},{"year":2004,"claim":"Identified the trafficking logic controlling surface activity, showing a C-terminal PDZ-binding motif recruits Mint-3 to recycle the enzyme to the membrane.","evidence":"Yeast two-hybrid, EWV deletion mutagenesis, Mint-3 siRNA, surface activity assays","pmids":["14990567"],"confidence":"High","gaps":["Endocytic route and other adaptors not mapped","Link to APP processing not yet examined"]},{"year":2009,"claim":"Connected MT5-MMP proteolysis to physiology by showing N-cadherin cleavage in nociceptors controls neuro-immune interactions and pain sensitivity.","evidence":"Mmp24 knockout mice, behavioral nociception assays, N-cadherin cleavage analysis","pmids":["19805319"],"confidence":"High","gaps":["Direct vs indirect N-cadherin cleavage in vivo not fully separated","Mast-cell interaction mechanism incompletely defined"]},{"year":2014,"claim":"Extended the N-cadherin-shedding function to adult neural stem cells, establishing a role in stem cell activation and quiescence.","evidence":"MT5-MMP knockout mice, N-cadherin shedding and NSC functional assays in vivo","pmids":["24952463"],"confidence":"High","gaps":["Downstream signaling from N-cadherin shedding not resolved","Dispensable for NSC identity—upstream regulators unknown"]},{"year":2015,"claim":"Defined MT5-MMP as an η-secretase that physically associates with APP and drives amyloidogenic processing independent of classical secretases.","evidence":"Reciprocal Co-IP and co-localization in HEKswe cells, MT5-MMP KO × 5xFAD cross, ELISA/Western","pmids":["26202697"],"confidence":"High","gaps":["Precise cleavage site generating sAPP95 not mapped here","Causal contribution to disease pathology not established"]},{"year":2017,"claim":"Linked the pro-amyloidogenic effect to subcellular trafficking by showing MT5-MMP localizes to early endosomes and routes APP/C99 there.","evidence":"Immunofluorescence, subcellular fractionation, Aβ40 ELISA in HEKswe cells","pmids":["28119565"],"confidence":"Medium","gaps":["Single cell model","Trafficking machinery directing endosomal sorting unidentified"]},{"year":2021,"claim":"Dissected proteolytic from non-proteolytic contributions, showing the C-terminal domains sort C99 to the endolysosomal system and protect it from proteasomal degradation even without catalytic activity.","evidence":"Domain deletion and catalytic-dead mutagenesis, fractionation, proteasome inhibitors, Western blot in HEKswe cells","pmids":["34117802"],"confidence":"High","gaps":["Direct C99-binding interface not structurally defined","Single cell system"]},{"year":2021,"claim":"Revealed transcriptional/post-transcriptional control in pain, with FTO-mediated m6A demethylation enhancing MMP24 translation to drive ERK-dependent hypersensitivity.","evidence":"MeRIP-seq, RIP, FTO overexpression/knockdown, ERK assays, neuropathic pain behavior","pmids":["33995105"],"confidence":"Medium","gaps":["Direct enzymatic substrate driving ERK activation unidentified","Single lab"]},{"year":2024,"claim":"Identified the activating protease, showing PCSK6 cleaves the RRRNKR propeptide motif to generate active MT5-MMP and enable amyloidogenic processing.","evidence":"Co-IP, cleavage-motif mutagenesis, PCSK6 knockdown in N2AAPP cells and AAV KD in APP23/PS45 mice","pmids":["38216110"],"confidence":"High","gaps":["Whether PCSK6 is the sole physiological activator unknown","Spatial site of activation not defined"]},{"year":2024,"claim":"Mapped organelle-dependence of η-secretase activity and identified C-terminal-domain binding partners that enhance sAPPη production.","evidence":"Subcellular-targeted fusion constructs with sAPPη ELISA; yeast two-hybrid screen of brain cDNA with functional follow-up","pmids":["38345408"],"confidence":"Medium","gaps":["Direct vs indirect nature of N4BP2L1/EIG121/BIN1/TMX3 effects not resolved","Endogenous relevance of nuclear/cytosolic localization unclear"]},{"year":2025,"claim":"Resolved the intracellular domain as a trafficking determinant, showing specific IC mutations divergently control C99 sorting, degradation, and Aβ output.","evidence":"IC-domain mutagenesis, high-content imaging, proximity ligation assay in human cell lines (preprint)","pmids":["bio_10.1101_2025.05.22.655451"],"confidence":"Medium","gaps":["Preprint, not yet peer-reviewed","Specific motifs/adaptors mediating each IC-cluster effect not defined"]},{"year":null,"claim":"How MT5-MMP's dual proteolytic and trafficking-scaffold functions are coordinated in vivo and whether targeting them alters Alzheimer pathology remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of the C-terminal/IC domain interactions with C99","Therapeutic modulation untested","Integration of pain, NSC, and amyloid roles in the same neurons unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,3,4,6,7,8]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0,3,4]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[10,14,15]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[2,5,13]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[9,13]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[13]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[13]}],"pathway":[{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[8,9,10]},{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[5,9,10,15]}],"complexes":[],"partners":["APP","MMP2","MINT-3","PCSK6","N4BP2L1","EIG121","BIN1","TMX3"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9Y5R2","full_name":"Matrix metalloproteinase-24","aliases":["Membrane-type matrix metalloproteinase 5","MT-MMP 5","MTMMP5","Membrane-type-5 matrix metalloproteinase","MT5-MMP","MT5MMP"],"length_aa":645,"mass_kda":73.2,"function":"Metalloprotease that mediates cleavage of N-cadherin (CDH2) and acts as a regulator of neuro-immune interactions and neural stem cell quiescence. Involved in cell-cell interactions between nociceptive neurites and mast cells, possibly by mediating cleavage of CDH2, thereby acting as a mediator of peripheral thermal nociception and inflammatory hyperalgesia. Key regulator of neural stem cells quiescence by mediating cleavage of CDH2, affecting CDH2-mediated anchorage of neural stem cells to ependymocytes in the adult subependymal zone, leading to modulate their quiescence. May play a role in axonal growth. Able to activate progelatinase A. May also be a proteoglycanase involved in degradation of proteoglycans, such as dermatan sulfate and chondroitin sulfate proteoglycans. Cleaves partially fibronectin, but not collagen type I, nor laminin (By similarity)","subcellular_location":"Secreted, extracellular space, extracellular matrix","url":"https://www.uniprot.org/uniprotkb/Q9Y5R2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MMP24","classification":"Not Classified","n_dependent_lines":8,"n_total_lines":1208,"dependency_fraction":0.006622516556291391},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/MMP24","total_profiled":1310},"omim":[{"mim_id":"604871","title":"MATRIX METALLOPROTEINASE 24; MMP24","url":"https://www.omim.org/entry/604871"},{"mim_id":"602285","title":"MATRIX METALLOPROTEINASE 17; MMP17","url":"https://www.omim.org/entry/602285"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoli","reliability":"Approved"},{"location":"Plasma membrane","reliability":"Approved"}],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"brain","ntpm":67.5}],"url":"https://www.proteinatlas.org/search/MMP24"},"hgnc":{"alias_symbol":["MT5-MMP"],"prev_symbol":[]},"alphafold":{"accession":"Q9Y5R2","domains":[{"cath_id":"1.10.101","chopping":"76-136","consensus_level":"high","plddt":74.6666,"start":76,"end":136},{"cath_id":"3.40.390.10","chopping":"168-327","consensus_level":"high","plddt":88.2574,"start":168,"end":327},{"cath_id":"2.110.10.10","chopping":"380-579","consensus_level":"high","plddt":86.8833,"start":380,"end":579}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y5R2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y5R2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y5R2-F1-predicted_aligned_error_v6.png","plddt_mean":73.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MMP24","jax_strain_url":"https://www.jax.org/strain/search?query=MMP24"},"sequence":{"accession":"Q9Y5R2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9Y5R2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9Y5R2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y5R2"}},"corpus_meta":[{"pmid":"10085137","id":"PMC_10085137","title":"Identification and characterization of the fifth membrane-type matrix metalloproteinase MT5-MMP.","date":"1999","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10085137","citation_count":215,"is_preprint":false},{"pmid":"10363975","id":"PMC_10363975","title":"Identification and characterization of human MT5-MMP, a new membrane-bound activator of progelatinase a overexpressed in brain tumors.","date":"1999","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/10363975","citation_count":202,"is_preprint":false},{"pmid":"24952463","id":"PMC_24952463","title":"MT5-MMP regulates adult neural stem cell functional quiescence through the cleavage of N-cadherin.","date":"2014","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/24952463","citation_count":88,"is_preprint":false},{"pmid":"26202697","id":"PMC_26202697","title":"MT5-MMP is a new pro-amyloidogenic proteinase that promotes amyloid pathology and cognitive decline in a transgenic 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potential impact on the survival of patients with non-small cell lung cancer (NSCLC).","date":"2021","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/35062057","citation_count":8,"is_preprint":false},{"pmid":"18062926","id":"PMC_18062926","title":"Identification of a novel human MT5-MMP transcript variant in multipotent NT2 cells.","date":"2007","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/18062926","citation_count":6,"is_preprint":false},{"pmid":"34359875","id":"PMC_34359875","title":"Deficiency in MT5-MMP Supports Branching of Human iPSCs-Derived Neurons and Reduces Expression of GLAST/S100 in iPSCs-Derived Astrocytes.","date":"2021","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/34359875","citation_count":5,"is_preprint":false},{"pmid":"19403421","id":"PMC_19403421","title":"[Inhibitory effects of RNA interference on MMP-24 expression and invasiveness of ovarian cancer SKOV(3) cells].","date":"2009","source":"Nan fang yi ke da xue xue bao = Journal of Southern Medical University","url":"https://pubmed.ncbi.nlm.nih.gov/19403421","citation_count":5,"is_preprint":false},{"pmid":"38216110","id":"PMC_38216110","title":"PCSK6 exacerbates Alzheimer's disease pathogenesis by promoting MT5-MMP maturation.","date":"2024","source":"Experimental neurology","url":"https://pubmed.ncbi.nlm.nih.gov/38216110","citation_count":4,"is_preprint":false},{"pmid":"39766352","id":"PMC_39766352","title":"Suppression of MT5-MMP Reveals Early Modulation of Alzheimer's Pathogenic Events in Primary Neuronal Cultures of 5xFAD Mice.","date":"2024","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/39766352","citation_count":2,"is_preprint":false},{"pmid":"40867559","id":"PMC_40867559","title":"Membrane-Type 5 Matrix Metalloproteinase (MT5-MMP): Background and Proposed Roles in Normal Physiology and Disease.","date":"2025","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/40867559","citation_count":1,"is_preprint":false},{"pmid":"38345408","id":"PMC_38345408","title":"Identification of binding partners that facilitate membrane-type 5 matrix metalloproteinase (MT5-MMP) processing of amyloid precursor protein.","date":"2024","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/38345408","citation_count":1,"is_preprint":false},{"pmid":"40330566","id":"PMC_40330566","title":"The Effect of Curcumin on the Activity of MMP-17 and MMP-24 in Hepatocytes of Mice Exposed to Thioacetamide.","date":"2024","source":"Reports of biochemistry & molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/40330566","citation_count":1,"is_preprint":false},{"pmid":"40178713","id":"PMC_40178713","title":"Downregulation of MT2-MMP and MT5-MMP in ulcerative colitis serves a diagnostic predictor and potential therapeutic targets.","date":"2025","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/40178713","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.05.22.655451","title":"Mutations in specific MT5-MMP domains prevent the accumulation of toxic APP metabolites and serve as templates for peptide-based therapeutics in cell models of Alzheimer’s disease","date":"2025-05-23","source":"bioRxiv","url":"https://doi.org/10.1101/2025.05.22.655451","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.07.26.24310191","title":"Disentangling shared genetic etiologies for kidney function and cardiovascular diseases","date":"2024-07-27","source":"bioRxiv","url":"https://doi.org/10.1101/2024.07.26.24310191","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":17074,"output_tokens":3688,"usd":0.053271,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11416,"output_tokens":3956,"usd":0.07799,"stage2_stop_reason":"end_turn"},"total_usd":0.131261,"stage1_batch_id":"msgbatch_01BbTCMyrRPyhtEq6b1MdwCN","stage2_batch_id":"msgbatch_011dEed5bbxYvTAaQWKJevxu","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1999,\n      \"finding\": \"MT5-MMP (MMP24) activates progelatinase A (MMP2) when co-expressed in MDCK cells, and this activity is dependent on proteolytic activity since a Glu→Ala mutation in the zinc-binding motif (HE255LGH) abolishes activation.\",\n      \"method\": \"Co-expression in MDCK cells, active-site mutagenesis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — active-site mutagenesis with functional readout, replicated in same year by independent lab (PMID:10363975)\",\n      \"pmids\": [\"10085137\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"MT5-MMP tends to shed from the cell surface as a soluble proteinase, distinguishing it from other MT-MMPs which remain membrane-bound.\",\n      \"method\": \"Cell expression and protein analysis (Western blot, conditioned media analysis)\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — demonstrated in cell expression system, consistent with independent replication\",\n      \"pmids\": [\"10085137\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Human MT5-MMP (MMP24) localizes to the plasma membrane when expressed in COS-7 cells, as shown by immunofluorescence and Western blot.\",\n      \"method\": \"Immunofluorescence and Western blot of transfected COS-7 cells\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — two orthogonal methods (IF + WB), single lab\",\n      \"pmids\": [\"10363975\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"The catalytic domain of MT5-MMP cleaves progelatinase A to generate the Mr 62,000 active form, demonstrated using a GST-fusion catalytic domain expressed in E. coli.\",\n      \"method\": \"In vitro proteolytic assay with recombinant catalytic domain (GST fusion)\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with purified recombinant protein, replicated in same year (PMID:10085137)\",\n      \"pmids\": [\"10363975\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Purified soluble MT5-MMP activates progelatinase A in a TIMP-2-sensitive fashion, and preferentially cleaves proteoglycans among extracellular matrix components.\",\n      \"method\": \"Biochemical purification, in vitro proteolytic assay with ECM substrate panel\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with purified enzyme and defined substrates, single lab\",\n      \"pmids\": [\"10622708\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"The C-terminal EWV motif of MT5-MMP serves as a retrieval signal for internalized MT5-MMP by interacting with the PDZ domain protein Mint-3, which recycles MT5-MMP to the plasma membrane. Deletion of the EWV signal impairs recycling and decreases surface activity. Mint-3 knockdown by siRNA decreases MT5-MMP activity.\",\n      \"method\": \"Yeast two-hybrid screening, deletion mutagenesis, siRNA knockdown, cell surface activity assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — yeast two-hybrid identification, mutagenesis of binding motif, siRNA KD, multiple orthogonal methods in one study\",\n      \"pmids\": [\"14990567\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"MT5-MMP cleaves N-cadherin in peptidergic nociceptors (CGRP+ dorsal root ganglion neurons); Mmp24-deficient mice show enhanced basal thermal sensitivity and failure to develop thermal hyperalgesia during inflammation, correlating with altered N-cadherin-mediated neuro-immune cell interactions.\",\n      \"method\": \"Mmp24 knockout mouse, behavioral nociception assays, N-cadherin cleavage analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with defined cellular and behavioral phenotype, mechanistic link to N-cadherin cleavage, multiple readouts\",\n      \"pmids\": [\"19805319\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MT5-MMP cleaves the N-cadherin ectodomain in adult neural stem cells (NSCs) and ependymocytes of the subependymal zone. MT5-MMP-mediated N-cadherin shedding is required for proper activation of NSCs under physiological and regenerative conditions but is dispensable for NSC generation and identity.\",\n      \"method\": \"MT5-MMP knockout mice, N-cadherin shedding assays, NSC functional assays in vivo\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO in vivo, specific substrate (N-cadherin) cleavage demonstrated, functional consequence on stem cell quiescence, rigorous controls\",\n      \"pmids\": [\"24952463\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"MT5-MMP co-localizes and co-immunoprecipitates with APP in HEKswe cells and promotes amyloidogenic APP processing, increasing levels of Aβ, C99, and a soluble APP fragment of 95 kDa (sAPP95). MT5-MMP deficiency in 5xFAD mice reduces Aβ, C99, and sAPP95 levels without altering α-, β-, or γ-secretase activities.\",\n      \"method\": \"Co-immunoprecipitation, co-localization (immunofluorescence), MT5-MMP KO × 5xFAD cross, ELISA, Western blot\",\n      \"journal\": \"Cellular and molecular life sciences : CMLS\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP in cell model, in vivo KO mouse validation, multiple orthogonal methods\",\n      \"pmids\": [\"26202697\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"MT5-MMP expressed in HEKswe cells localizes to early endosomes and promotes trafficking of APP and C99 into early endosomes, as well as increased Aβ40 in cell supernatants, establishing a pro-amyloidogenic role via endosomal trafficking.\",\n      \"method\": \"Immunofluorescence co-localization, subcellular fractionation, ELISA in transfected HEKswe cells\",\n      \"journal\": \"Frontiers in molecular neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — subcellular localization linked to functional APP trafficking consequence, single lab\",\n      \"pmids\": [\"28119565\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Deletion of C-terminal non-catalytic domains of MT5-MMP hampers its ability to process APP and release sAPP95. Catalytically inactive MT5-MMP variants still increase Aβ and promote APP/C99 sorting to the endolysosomal system via interactions of the C-terminal portion with C99. Deletion of the C-terminal domain causes proteasomal degradation of C99 and prevents Aβ accumulation.\",\n      \"method\": \"Domain deletion mutagenesis, catalytic-dead variants, subcellular fractionation, proteasome inhibitors, Western blot in HEKswe cells\",\n      \"journal\": \"FASEB journal : official publication of the Federation of American Societies for Experimental Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — structure-function mutagenesis with multiple deletion variants, both proteolytic and non-proteolytic mechanisms dissected, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"34117802\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"FTO (m6A eraser) binds to Mmp24 mRNA in spinal cord neurons after spinal nerve ligation, reducing m6A modification and facilitating MMP24 translation. MMP24 upregulation in spinal cord neurons promotes ERK activation and nociceptive hypersensitivity.\",\n      \"method\": \"MeRIP-seq, RNA immunoprecipitation, FTO overexpression/knockdown in spinal neurons, ERK activation assays, neuropathic pain behavioral tests\",\n      \"journal\": \"Frontiers in pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MeRIP-seq + RIP assay + FTO OE/KD, multiple methods, single lab\",\n      \"pmids\": [\"33995105\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PCSK6 cleaves and activates MT5-MMP by recognizing the RRRNKR sequence in the MT5-MMP N-terminal propeptide domain. Mutation or knockout of this cleavage motif prevents PCSK6 from interacting with and cleaving MT5-MMP, reducing amyloidogenic APP processing.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis of cleavage motif, PCSK6 knockdown in N2AAPP cells and AAV-mediated KD in APP23/PS45 mice\",\n      \"journal\": \"Experimental neurology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — cleavage motif mutagenesis abolishes interaction, in vitro cell and in vivo AAV KD validation, multiple methods\",\n      \"pmids\": [\"38216110\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"MT5-MMP localizes to the nucleus, cytosol, and cytosolic subcellular granules in CHO cells stably expressing APP751. Using subcellular-targeted MT5-MMP fusion proteins, η-secretase (sAPPη) activity was highest when MT5-MMP was directed to the endosome, Golgi, plasma membrane, or mitochondria.\",\n      \"method\": \"Subcellular-targeted fusion protein constructs, sAPPη ELISA, immunofluorescence localization in CHO-APP751 cells\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — targeted fusion proteins to specific organelles with functional readout, single lab\",\n      \"pmids\": [\"38345408\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Yeast two-hybrid screening identified N4BP2L1, TMX3, EIG121, BIN1, RUFY4, HTRA1, and TMEM199 as binding partners of the MT5-MMP C-terminal domain in a human brain cDNA library. Binding of N4BP2L1, EIG121, BIN1, or TMX3 resulted in significant increase in sAPPη production.\",\n      \"method\": \"Yeast two-hybrid screen, co-expression functional assay (sAPPη ELISA)\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast two-hybrid identification with functional follow-up in cell model, single lab\",\n      \"pmids\": [\"38345408\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Specific amino acid mutations in the intracellular (IC) domain of MT5-MMP induce C99 degradation and decrease Aβ levels, while mutations in different IC clusters have divergent effects. The IC domain regulates subcellular trafficking of C99 through the endomembrane system and proximity with C99 as shown by proximity ligation assay.\",\n      \"method\": \"Domain deletion/substitution mutagenesis, high-content imaging, proximity ligation assay, co-transfection in human cell lines\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — mutagenesis with functional readouts and PLA, preprint not yet peer-reviewed\",\n      \"pmids\": [\"bio_10.1101_2025.05.22.655451\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"MT5-MMP (MMP24) is a brain-enriched membrane-type matrix metalloproteinase that acts as an η-secretase cleaving APP to generate pro-amyloidogenic fragments (Aβ, C99, sAPP95); it is activated by PCSK6-mediated cleavage of its RRRNKR propeptide motif, traffics to the cell surface via a C-terminal EWV motif interaction with the PDZ adaptor Mint-3, cleaves N-cadherin to regulate neural stem cell quiescence and nociceptor-mast cell interactions, degrades proteoglycans, and activates progelatinase A (MMP2) through its catalytic zinc-binding domain, with its non-catalytic C-terminal domains additionally controlling APP/C99 sorting through the endolysosomal system via interactions with binding partners including N4BP2L1, EIG121, BIN1, and TMX3.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"MMP24 (MT5-MMP) is a brain-enriched membrane-type matrix metalloproteinase that couples extracellular and intracellular proteolytic functions to neural stem cell regulation, nociception, and amyloidogenic APP processing [#6, #7, #8]. Its catalytic zinc-binding domain activates progelatinase A (MMP2) and preferentially degrades proteoglycans among extracellular matrix components, an activity abolished by mutation of the HEXGH zinc-binding motif and sensitive to TIMP-2 [#0, #3, #4]. Surface presentation of the enzyme is controlled by recycling: its C-terminal EWV motif binds the PDZ adaptor Mint-3 to retrieve internalized MT5-MMP back to the plasma membrane, and loss of this signal or Mint-3 depletion reduces surface activity [#5]. Activation requires PCSK6, which recognizes and cleaves the RRRNKR propeptide motif to generate active enzyme [#12]. In neurons, MT5-MMP sheds the N-cadherin ectodomain to regulate adult neural stem cell activation and quiescence and to modulate nociceptor neuro-immune interactions, with knockout mice showing altered thermal sensitivity and impaired stem cell activation [#6, #7]. Acting as an \\u03b7-secretase, MT5-MMP associates with APP and promotes amyloidogenic processing, raising A\\u03b2, C99, and sAPP95 without altering \\u03b1-, \\u03b2-, or \\u03b3-secretase activities, in part by directing APP and C99 into the endolysosomal system [#8, #9, #10]. This pro-amyloidogenic activity is only partly proteolytic: the C-terminal non-catalytic and intracellular domains govern C99 sorting and stability through brain-derived binding partners including N4BP2L1, EIG121, BIN1, and TMX3, and deletion of these domains routes C99 to proteasomal degradation [#10, #14, #15].\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"Established MT5-MMP as a catalytically active metalloproteinase capable of activating a downstream proteinase, defining its core enzymatic function and zinc-dependent mechanism.\",\n      \"evidence\": \"Co-expression in MDCK cells with active-site (Glu\\u2192Ala) mutagenesis, plus in vitro assays with recombinant catalytic domain and purified soluble enzyme\",\n      \"pmids\": [\"10085137\", \"10363975\", \"10622708\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological MMP2 activation in vivo not demonstrated\", \"Full ECM substrate repertoire beyond proteoglycans not defined\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Localized MT5-MMP to the plasma membrane and showed it can shed as a soluble proteinase, distinguishing it from membrane-retained MT-MMPs.\",\n      \"evidence\": \"Immunofluorescence and Western blot of transfected COS-7 and conditioned-media analysis\",\n      \"pmids\": [\"10363975\", \"10085137\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Shedding mechanism and protease responsible not identified\", \"Relative balance of membrane vs soluble pools in neurons unknown\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Identified the trafficking logic controlling surface activity, showing a C-terminal PDZ-binding motif recruits Mint-3 to recycle the enzyme to the membrane.\",\n      \"evidence\": \"Yeast two-hybrid, EWV deletion mutagenesis, Mint-3 siRNA, surface activity assays\",\n      \"pmids\": [\"14990567\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Endocytic route and other adaptors not mapped\", \"Link to APP processing not yet examined\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Connected MT5-MMP proteolysis to physiology by showing N-cadherin cleavage in nociceptors controls neuro-immune interactions and pain sensitivity.\",\n      \"evidence\": \"Mmp24 knockout mice, behavioral nociception assays, N-cadherin cleavage analysis\",\n      \"pmids\": [\"19805319\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct vs indirect N-cadherin cleavage in vivo not fully separated\", \"Mast-cell interaction mechanism incompletely defined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Extended the N-cadherin-shedding function to adult neural stem cells, establishing a role in stem cell activation and quiescence.\",\n      \"evidence\": \"MT5-MMP knockout mice, N-cadherin shedding and NSC functional assays in vivo\",\n      \"pmids\": [\"24952463\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Downstream signaling from N-cadherin shedding not resolved\", \"Dispensable for NSC identity\\u2014upstream regulators unknown\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Defined MT5-MMP as an \\u03b7-secretase that physically associates with APP and drives amyloidogenic processing independent of classical secretases.\",\n      \"evidence\": \"Reciprocal Co-IP and co-localization in HEKswe cells, MT5-MMP KO \\u00d7 5xFAD cross, ELISA/Western\",\n      \"pmids\": [\"26202697\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Precise cleavage site generating sAPP95 not mapped here\", \"Causal contribution to disease pathology not established\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Linked the pro-amyloidogenic effect to subcellular trafficking by showing MT5-MMP localizes to early endosomes and routes APP/C99 there.\",\n      \"evidence\": \"Immunofluorescence, subcellular fractionation, A\\u03b240 ELISA in HEKswe cells\",\n      \"pmids\": [\"28119565\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single cell model\", \"Trafficking machinery directing endosomal sorting unidentified\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Dissected proteolytic from non-proteolytic contributions, showing the C-terminal domains sort C99 to the endolysosomal system and protect it from proteasomal degradation even without catalytic activity.\",\n      \"evidence\": \"Domain deletion and catalytic-dead mutagenesis, fractionation, proteasome inhibitors, Western blot in HEKswe cells\",\n      \"pmids\": [\"34117802\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct C99-binding interface not structurally defined\", \"Single cell system\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Revealed transcriptional/post-transcriptional control in pain, with FTO-mediated m6A demethylation enhancing MMP24 translation to drive ERK-dependent hypersensitivity.\",\n      \"evidence\": \"MeRIP-seq, RIP, FTO overexpression/knockdown, ERK assays, neuropathic pain behavior\",\n      \"pmids\": [\"33995105\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct enzymatic substrate driving ERK activation unidentified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified the activating protease, showing PCSK6 cleaves the RRRNKR propeptide motif to generate active MT5-MMP and enable amyloidogenic processing.\",\n      \"evidence\": \"Co-IP, cleavage-motif mutagenesis, PCSK6 knockdown in N2AAPP cells and AAV KD in APP23/PS45 mice\",\n      \"pmids\": [\"38216110\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether PCSK6 is the sole physiological activator unknown\", \"Spatial site of activation not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Mapped organelle-dependence of \\u03b7-secretase activity and identified C-terminal-domain binding partners that enhance sAPP\\u03b7 production.\",\n      \"evidence\": \"Subcellular-targeted fusion constructs with sAPP\\u03b7 ELISA; yeast two-hybrid screen of brain cDNA with functional follow-up\",\n      \"pmids\": [\"38345408\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect nature of N4BP2L1/EIG121/BIN1/TMX3 effects not resolved\", \"Endogenous relevance of nuclear/cytosolic localization unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Resolved the intracellular domain as a trafficking determinant, showing specific IC mutations divergently control C99 sorting, degradation, and A\\u03b2 output.\",\n      \"evidence\": \"IC-domain mutagenesis, high-content imaging, proximity ligation assay in human cell lines (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.05.22.655451\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, not yet peer-reviewed\", \"Specific motifs/adaptors mediating each IC-cluster effect not defined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How MT5-MMP's dual proteolytic and trafficking-scaffold functions are coordinated in vivo and whether targeting them alters Alzheimer pathology remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of the C-terminal/IC domain interactions with C99\", \"Therapeutic modulation untested\", \"Integration of pain, NSC, and amyloid roles in the same neurons unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 3, 4, 6, 7, 8]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0, 3, 4]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [10, 14, 15]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [2, 5, 13]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [9, 13]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [13]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [13]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:1474244\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [8, 9, 10]},\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [5, 9, 10, 15]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"APP\", \"MMP2\", \"Mint-3\", \"PCSK6\", \"N4BP2L1\", \"EIG121\", \"BIN1\", \"TMX3\"],\n    \"other_free_text\": []\n  }\n}\n```","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":7,"faith_pct":85.71428571428571}}