{"gene":"ADAMTS4","run_date":"2026-06-09T22:02:41","timeline":{"discoveries":[{"year":2000,"finding":"ADAMTS4 (aggrecanase-1) cleaves aggrecan at the Glu373-Ala374 bond in the interglobular domain and at four additional sites within the chondroitin sulfate-rich region (KEEE1667-1668GLGS, GELE1480-1481GRGT, TAQE1771-1772AGEG, VSQE1871-1872LGQR), with cleavage in the CS-rich region occurring more efficiently than at the IGD site.","method":"In vitro cleavage assay with recombinant human ADAMTS4 on native aggrecan, followed by N-terminal sequence analysis of fragments","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct in vitro enzymatic assay with sequence-confirmed cleavage sites, replicated across multiple studies","pmids":["10751421"],"is_preprint":false},{"year":2000,"finding":"The thrombospondin type-1 (TSP-1) motif of ADAMTS4 binds to glycosaminoglycans of aggrecan and is required for aggrecan substrate recognition and cleavage; truncated ADAMTS4 lacking the TSP-1 motif failed to cleave aggrecan, TSP-1 peptides blocked cleavage by competing with enzyme binding, and GAG-free aggrecan was not cleaved.","method":"Domain truncation mutagenesis, peptide competition assays, aggrecan cleavage assays with GAG-free aggrecan","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — multiple orthogonal in vitro approaches including mutagenesis and competition assays demonstrating substrate binding mechanism","pmids":["10827174"],"is_preprint":false},{"year":2000,"finding":"ADAMTS4 cleaves brevican at the Glu395-Ser396 bond within the central non-homologous domain, a different site from that cleaved by MMPs (Ala360-Phe361), demonstrating substrate specificity distinct from MMP family members.","method":"In vitro digestion of brevican with purified ADAMTS4 and MMPs, followed by N-terminal sequence analysis of cleavage fragments","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct in vitro biochemical assay with sequence-confirmed cleavage sites","pmids":["10986281"],"is_preprint":false},{"year":2001,"finding":"ADAMTS4 cleaves versican V1 at the Glu441-Ala442 bond, generating the DPEAAE neoepitope, as demonstrated by recombinant enzyme cleavage of recombinant and native human versican substrates; mature ADAMTS4 protein was also detected in aortic intima extracts.","method":"In vitro cleavage assay with recombinant ADAMTS4 and recombinant/native versican V1 substrates; Western analysis with neoepitope antisera; Western analysis of aortic tissue","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct in vitro enzymatic assay with sequence-confirmed cleavage site, combined with tissue detection","pmids":["11278559"],"is_preprint":false},{"year":2001,"finding":"TIMP-3 is a potent inhibitor of ADAMTS4 (aggrecanase-1) with Ki values in the subnanomolar range; the N-terminal inhibitory domain of TIMP-3 alone is sufficient for this inhibition, whereas TIMP-1, TIMP-2 and TIMP-4 do not effectively inhibit ADAMTS4.","method":"In vitro inhibition kinetics assay with N-terminal TIMP-3 domain expressed from bacterial inclusion bodies","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — quantitative in vitro inhibition assay with recombinant proteins, replicated across multiple studies","pmids":["11278243"],"is_preprint":false},{"year":2002,"finding":"ADAMTS4 activation requires C-terminal truncation from the p75 (or p68) form to the p60/p53 forms; only the p60 and p53 forms exhibit aggrecanase and versicanase activity; this truncation is mediated by a MMP (GPI-anchored type) and blocked by TIMP-1, MMP inhibitors, furin inhibitors, and inhibitors of GPI synthesis, but not by serine or cysteine protease inhibitors.","method":"Stable transfection of human chondrosarcoma cells with full-length ADAMTS4; domain-specific antisera Western analysis; preparative SDS-PAGE isolation of individual forms with activity assays; pharmacological inhibition studies","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods including fractionation, activity assays, and pharmacological dissection; replicated by other studies","pmids":["11796708"],"is_preprint":false},{"year":2002,"finding":"Full-length ADAMTS4 undergoes autocatalytic C-terminal truncation to generate ~53 kDa and ~40 kDa isoforms with reduced affinity for sulfated GAGs; the cysteine-rich and spacer domains contain additional GAG-binding sites beyond the thrombospondin motif, as demonstrated by competition with synthetic peptides mimicking GAG-binding consensus sequences.","method":"C-terminal sequencing and mass analysis of autocatalytic fragments; GAG-binding competition assays with deglycosylated aggrecan and synthetic peptides","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct biochemical characterization with sequence analysis and binding competition experiments","pmids":["12202483"],"is_preprint":false},{"year":2002,"finding":"ADAMTS4 also cleaves the Asn341-Phe342 MMP site in aggrecan interglobular domain (secondarily to the Glu373-Ala374 aggrecanase site); this secondary cleavage is inhibited by TIMP-3 but not TIMP-1 or TIMP-2, distinguishing it from MMP activity.","method":"In vitro digestion of native human aggrecan and recombinant G1-G2 constructs with purified ADAMTS4; inhibitor studies with TIMP-1, TIMP-2, TIMP-3","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro enzymatic assay with mutagenesis of cleavage sites and selective inhibitor validation","pmids":["11854269"],"is_preprint":false},{"year":2003,"finding":"The non-catalytic C-terminal spacer domain of ADAMTS4 masks its general proteolytic activity; deletion of the spacer domain generates a more promiscuous enzyme with enhanced activity against the Glu373-Ala374 IGD bond, Cm-Tf, fibromodulin, and decorin, while removal of the cysteine-rich domain reduces aggrecanase activity by 80%; the full-length 70 kDa form is the most effective aggrecanase but shows little IGD activity and binds to the pericellular matrix, whereas spacer-deleted ADAMTS4 is released from the matrix.","method":"Expression of domain deletion mutants in mammalian cells; aggrecanase activity assays; general proteinase substrate (Cm-Tf) assays; Western analysis with domain-specific antisera","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — systematic domain deletion mutagenesis with multiple substrate activity assays and localization studies","pmids":["14662755"],"is_preprint":false},{"year":2003,"finding":"Proprotein convertase furin cleaves the prodomain of pro-ADAMTS4 in the trans-Golgi network at multiple sites (RPRR206-209, RAKR209-212, or KR211-212), and the pro-form (but not the mature form) physically co-precipitates with furin; processing is blocked by brefeldin A and furin-specific inhibitors; a furin-independent activation pathway also exists.","method":"Co-localization by confocal microscopy; co-immunoprecipitation; RNA interference of furin; furin-specific inhibitor treatment; brefeldin A treatment","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-precipitation, RNAi, pharmacological inhibition, and localization in a single study","pmids":["14744861"],"is_preprint":false},{"year":2003,"finding":"C-terminal truncation of ADAMTS4 from p68 to p53 (required for activation of IGD cleavage) is mediated by GPI-anchored MT4-MMP (MMP-17) on the cell surface; co-transfection with active MT4-MMP enhanced truncation, while inactive MT4-MMP mutant was ineffective; activated p53 ADAMTS4 remains associated with cell surface syndecan-1 through both chondroitin sulfate and heparan sulfate chains.","method":"Co-transfection of ADAMTS4 with active/inactive MT4-MMP; phosphatidylinositol-specific phospholipase C treatment; glycosaminoglycan lyase digestions with FACE analysis; immunoprecipitation with anti-syndecan-1","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods including active vs. inactive mutant controls, enzymatic digestion, and co-precipitation; independently supported by other studies on MT4-MMP/ADAMTS4 axis","pmids":["14701864"],"is_preprint":false},{"year":2004,"finding":"ADAMTS4 binds to the C-terminal domain of fibronectin through its spacer domain; this interaction inhibits aggrecanase activity of ADAMTS4 (IC50 ~110 nM for full-length fibronectin; ~170 nM for 40-kDa C-terminal fibronectin fragment); ADAMTS4 lacking the spacer domain is not inhibited by fibronectin and does not co-localize with fibronectin on cell surfaces.","method":"Yeast two-hybrid screening; chemical cross-linking; solid-phase binding assay; confocal microscopy co-localization; aggrecanase activity assay with inhibition by fibronectin","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — yeast two-hybrid, cross-linking, solid-phase binding, and functional inhibition assay in a single study with domain mapping","pmids":["15161923"],"is_preprint":false},{"year":2004,"finding":"ADAMTS4 cleaves versican V2 at Glu405-Gln406 to generate the brain protein GHAP (glial hyaluronate binding protein), as demonstrated by immunological analysis of purified human GHAP with anti-neoepitope antiserum and in vitro digestion of human cerebellum proteoglycans with ADAMTS4.","method":"Anti-neoepitope antiserum immunological analysis; in vitro digestion of human cerebellum proteoglycans with ADAMTS4; Western analysis","journal":"The Biochemical journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — biochemical identification with neoepitope antibody and in vitro digestion, single lab","pmids":["14561220"],"is_preprint":false},{"year":2005,"finding":"Pro-ADAMTS4 is not capable of autocatalytic prodomain removal under physiological conditions (mercuric compounds, temperatures 22-65°C, ionic strengths 0.1-1M, neutral/acidic pH do not trigger autocatalysis); proprotein convertases furin, PACE4, and PC5/6 efficiently remove the prodomain at Arg212/Phe213 to generate active enzyme; MMP-9 and trypsin can also remove the prodomain; autocatalysis occurs only at basic pH 8-10.","method":"In vitro incubation of pro-ADAMTS4 with diverse proteases and conditions; activity assays; N-terminal sequencing","journal":"Archives of biochemistry and biophysics","confidence":"High","confidence_rationale":"Tier 1 / Moderate — comprehensive in vitro assay screening multiple conditions with activity readout, single lab but systematic approach","pmids":["16289022"],"is_preprint":false},{"year":2005,"finding":"MT4-MMP plays a central role in IL-1-induced cartilage aggrecanolysis by activating ADAMTS4; IL-1 treatment causes loss of p68 ADAMTS4 from cartilage and appearance of p53 in medium; blocking MT4-MMP (via ManN or EST) nearly completely prevented p68 loss, p53 formation, and aggrecan degradation; increased ADAMTS4/5 mRNA by IL-1 was not accompanied by increased total enzyme protein, indicating that activation rather than new synthesis drives aggrecanolysis.","method":"Western analysis of aggrecan fragments and ADAMTS4 species in cartilage explants; real-time PCR; pharmacological inhibition with ManN (GPI synthesis inhibitor) and EST (MMP expression inhibitor)","journal":"Osteoarthritis and cartilage","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple Western analyses and pharmacological inhibition in explant system, single lab","pmids":["15780640"],"is_preprint":false},{"year":2006,"finding":"The spacer domain of ADAMTS4 is critical for its localization in the extracellular matrix, whereas for ADAMTS5 the cysteine-rich domain is more influential; sequential inclusion of C-terminal ancillary domains of ADAMTS4 progressively enhances activity against aggrecan, Cm-Tf, fibromodulin, decorin, biglycan, and fibronectin; very little proteolytic activity is detected with the catalytic domain alone.","method":"Expression of domain deletion mutants; activity assays against multiple substrates; ECM localization assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — systematic domain deletion mutagenesis with multiple substrate assays, comparative study with ADAMTS5 providing internal controls","pmids":["17430884"],"is_preprint":false},{"year":2006,"finding":"The NFATp and Runx2 transcription factors regulate ADAMTS4 promoter activity in chondrocytes; the NFI binding site (-441 to -429) acts as a negative regulator of the ADAMTS4 promoter specifically in chondrocytes; the region -383 to +10 is necessary for full basal promoter activity and contains Sp1 and AP2 sites.","method":"Reporter gene (promoter-luciferase) assays with deletion variants; mRNA analysis in porcine chondrocytes and NIH3T3 cells","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — promoter reporter assays with deletion constructs in relevant cell types, single lab","pmids":["16677612","11254106"],"is_preprint":false},{"year":2007,"finding":"TIMP-3 inhibition of ADAMTS4 is enhanced in the presence of aggrecan through binding of chondroitin 6-sulfate GAGs to the thrombospondin type 1 motif and spacer domains of ADAMTS4, forming a complex with improved TIMP-3 binding affinity over free ADAMTS4.","method":"FRET peptide assay; aggrecan-binding competition experiments; solid-phase binding assays","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — quantitative FRET assay and binding studies, single lab, mechanistic dissection of substrate-inhibitor interplay","pmids":["17470431"],"is_preprint":false},{"year":2007,"finding":"Crystal structures of ADAMTS4 reveal two distinct catalytic site configurations: a closed autoinhibited non-binding form and an open binding-competent form; mature aggrecanases exist as an ensemble of at least two isomers, only one of which is proteolytically active.","method":"X-ray crystallography of human ADAMTS4 in apo and inhibitor-bound forms","journal":"Protein science","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure determination, single study but direct structural evidence","pmids":["18042673"],"is_preprint":false},{"year":2007,"finding":"Aggrecan cleavage by ADAMTS4 proceeds via an exosite mechanism: substrate initially binds at an exosite (reflected in the apparent Km), then the peptide sequence binds at the active site; an active-site inhibitor (SC81956, hydroxamic acid, Ki=23 nM) is non-competitive with aggrecan but competitive with low-molecular-weight peptide substrates, consistent with inability to compete with the final Michaelis complex.","method":"Enzyme kinetics with native aggrecan and fluorogenic peptide substrates; inhibition kinetics with varying substrate concentrations","journal":"Biochemistry","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — rigorous kinetic analysis with two substrate types, single lab","pmids":["17487981"],"is_preprint":false},{"year":2008,"finding":"Calcium pentosan polysulfate (CaPPS) directly inhibits the aggrecanase activity of ADAMTS4 by interacting with its C-terminal ancillary domains (thrombospondin type 1 repeat, cysteine-rich, and spacer domains), without affecting mRNA expression of ADAMTS species.","method":"Aggrecanase activity assay in IL-1α-stimulated OA chondrocytes; synthetic peptide competition of CaPPS binding; mRNA expression analysis","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct activity assay and domain mapping by peptide competition, single lab","pmids":["18671975"],"is_preprint":false},{"year":2009,"finding":"ADAMTS4 processes the matricellular protein hevin in mouse brain; in vitro digestion of hevin with ADAMTS4 produced fragments similar to those present in brain lysates; a SPARC-like fragment generated from hevin co-localizes with ADAMTS4 in vivo in mouse cerebellum, and this proteolysis contributes to normal cerebellar development.","method":"In vitro digestion of hevin with recombinant ADAMTS4; co-localization by confocal microscopy with monoclonal antibodies; comparison of brain lysate fragments","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro cleavage assay combined with in vivo co-localization, single lab","pmids":["20018883"],"is_preprint":false},{"year":2012,"finding":"ADAMTS4 is endocytosed and degraded by chondrocytes via LRP1; the cysteine-rich and spacer domains of ADAMTS4 are responsible for binding to LRP1 clusters II and IV (KD,app ~98 nM and ~73 nM, respectively); the half-life of ADAMTS4 endocytosis is ~220 min, slower than ADAMTS5 (~100 min) due to 13-fold lower affinity for LRP1; ADAMTS5 competitively inhibits ADAMTS4 endocytosis but not vice versa.","method":"Domain deletion mutagenesis; soluble LRP1 cluster binding assays; endocytosis kinetics; competitive inhibition assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — domain mutagenesis, quantitative binding affinity measurements, kinetic endocytosis assays, with clear mechanistic dissection","pmids":["24474687"],"is_preprint":false},{"year":2012,"finding":"ADAMTS4 degrades brevican, neurocan, and phosphacan core proteins and reverses their inhibition of neurite outgrowth; local administration of ADAMTS4 protein promoted motor function recovery and enhanced axonal regeneration/sprouting after spinal cord contusion injury in mice.","method":"In vitro CSPG degradation assay; neurite outgrowth assay; in vivo spinal cord contusion model with local ADAMTS4 administration and behavioral assessment","journal":"Journal of neuroinflammation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro substrate degradation combined with in vivo functional assay, single lab","pmids":["22420304"],"is_preprint":false},{"year":2012,"finding":"ADAMTS4 and ADAMTS5 participate in regulated proteolytic processing of Reelin; both enzymes cleave Reelin in vitro, and TIMP-3, α-2-Macroglobulin, serpins, and MMP-9 modulate this cleavage; ADAMTS4 and Reelin expression levels largely overlap in murine hippocampus.","method":"In vitro cleavage assay with recombinant enzymes and Reelin; inhibitor studies with TIMPs and serpins; immunolocalization in murine hippocampus","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct in vitro cleavage assay with inhibitor dissection, combined with in vivo localization, single lab","pmids":["23082219"],"is_preprint":false},{"year":2013,"finding":"TNF-α and IL-1β regulate ADAMTS4 expression in nucleus pulposus cells through MAPK (ERK1, p38α, p38β2, p38γ) and NF-κB signaling; p65 induces ADAMTS4 promoter activity while p50 blocks it; silencing of NF-κB components (p65, p52, IKK-α, IKK-β) decreased ADAMTS-4 and -5 levels and aggrecan degradation; both ADAMTS-4 and ADAMTS-5 contribute non-redundantly to aggrecan degradation in human NP cells.","method":"Transient transfection promoter assays; MAPK/NF-κB inhibitors; gain/loss-of-function studies; lentiviral shRNA silencing; Western blotting; aggrecan degradation assays","journal":"The American journal of pathology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple pathway inhibitors and genetic knockdown with functional readout, single lab","pmids":["23602832"],"is_preprint":false},{"year":2013,"finding":"IL-1β-induced upregulation of ADAMTS4 in human OA chondrocytes is suppressed by miR-125b overexpression; luciferase reporter assay with mutated miR-125b binding site in the ADAMTS4 3'UTR confirmed direct interaction, establishing miR-125b as a direct post-transcriptional regulator of ADAMTS4.","method":"miR-125b overexpression; luciferase reporter assay with wild-type and mutant ADAMTS4 3'UTR; quantitative RT-PCR; Western blotting","journal":"Arthritis research & therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — luciferase assay with mutation validation plus functional overexpression experiment, single lab","pmids":["23406982"],"is_preprint":false},{"year":2015,"finding":"CCN1 (Cyr61) binds specifically to the cysteine-rich domain of ADAMTS4 and inhibits its aggrecanase activity; CCN1 co-purifies with ADAMTS4-transfected chondrocytic cells (identified by LC-MS/MS), and the interaction was confirmed by immunoprecipitation and solid-phase binding assay; in TGFβ-treated chondrocytes, CCN1 knockdown reveals latent ADAMTS4 aggrecanase activity.","method":"LC-MS/MS identification of co-purified proteins; immunoprecipitation; solid-phase binding assay; aggrecan digestion assay; siRNA knockdown; immunohistochemistry","journal":"Arthritis & rheumatology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple binding assays combined with functional inhibition and genetic knockdown, single lab","pmids":["25709087"],"is_preprint":false},{"year":2017,"finding":"ADAMTS4 translocates to the nucleus in smooth muscle cells (SMCs) under stress and directly cleaves and degrades poly ADP ribose polymerase-1 (PARP-1), leading to SMC apoptosis; ADAMTS4 deficiency in mice significantly reduced angiotensin II/high-fat diet-induced aortic aneurysm formation, elastic fiber destruction, versican degradation, macrophage infiltration, and apoptosis.","method":"Adamts4-/- mouse model of sporadic AAD (angiotensin II + high fat diet); nuclear translocation demonstrated by imaging of apoptotic SMCs; direct PARP-1 cleavage assay; aortic phenotyping (diameter, histology)","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — knockout mouse model combined with direct cleavage assay, single lab; nuclear localization is notable but mechanistic evidence for PARP-1 cleavage relies on single study","pmids":["28955046"],"is_preprint":false},{"year":2018,"finding":"ADAMTS4 cleaves APP/Aβ to generate N-truncated Aβ4-x peptides; a recognition site for ADAMTS4 was identified in the Aβ sequence; inducible overexpression of ADAMTS4 in HEK293 cells increased secretion of Aβ4-40; in ADAMTS4-/- mice (5xFAD background) Aβ4-40 levels were reduced; ADAMTS4 is exclusively expressed in oligodendrocytes in adult mouse brain, and ADAMTS4-/- oligodendrocyte cultures do not produce Aβ4-40.","method":"ADAMTS4 recognition site identification; inducible overexpression in HEK293 cells with Aβ ELISA; ADAMTS4-/- mouse model; primary oligodendrocyte cultures from ADAMTS4-/- mice; immunofluorescence co-localization","journal":"Acta neuropathologica","confidence":"High","confidence_rationale":"Tier 2 / Strong — overexpression gain-of-function, knockout loss-of-function, and cell-specific culture experiments with consistent results, single lab but multiple orthogonal approaches","pmids":["30426203"],"is_preprint":false},{"year":2018,"finding":"Sox4 (and Sox11) directly bind to the ADAMTS4 promoter and upregulate ADAMTS4 and ADAMTS5 gene expression; demonstrated by luciferase reporter assay and chromatin immunoprecipitation; Sox4 overexpression in mouse femoral head cartilage organ cultures caused cartilage destruction associated with increased aggrecanase expression.","method":"Luciferase reporter assay; chromatin immunoprecipitation (ChIP); adenoviral overexpression in mouse femoral head organ cultures; quantitative RT-PCR","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and reporter assay confirm direct promoter binding with functional validation in organ culture, single lab","pmids":["30016600"],"is_preprint":false},{"year":2020,"finding":"Damage-responsive lung fibroblasts produce ADAMTS4 during severe respiratory viral infection; ADAMTS4 ECM protease activity modifies the lung microenvironment to promote immune cell infiltration; ADAMTS4 levels in lower respiratory tract samples correlated with severity of influenza infection in three human cohorts.","method":"Single-cell transcriptomics identifying fibroblast activation states; mouse influenza model with fibroblast characterization; measurement of ADAMTS4 in human BALF from influenza patients","journal":"Nature","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mechanistic identification of ADAMTS4-producing fibroblast subpopulation with human cohort validation, but specific molecular mechanism of immune cell infiltration promotion is correlative","pmids":["33116313"],"is_preprint":false},{"year":2021,"finding":"ADAMTS4 (along with ADAMTS1 and ADAMTS5) cleaves versican V1 at multiple sites beyond the known Glu441-Ala442 bond; 21 novel cleavage sites were identified using LC-MS/MS label-free proteomics with z-score ranking; ADAMTS4 shows a substrate site preference for P1-Glu residue.","method":"In vitro digestion of recombinant versican V1 with recombinant full-length ADAMTS4 vs. catalytically inactive mutant; LC-MS/MS label-free quantitative proteomics with z-score analysis","journal":"Journal of proteomics","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — systematic in vitro enzymatic assay with inactive mutant controls and quantitative proteomics, single lab","pmids":["34450332"],"is_preprint":false},{"year":2011,"finding":"Adamts4 and Adamts1 play redundant roles in perinatal kidney development; Adamts4 single-knockout mice are phenotypically normal, but >95% of Adamts1-/-;Adamts4-/- double-knockout mice die within 72 hours of birth with marked thinning of the renal medulla, a defect not observed in embryos.","method":"Gene targeting to generate Adamts4-/- mice; genetic cross with Adamts1-/- mice; histological analysis of renal medulla","journal":"Developmental dynamics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis using double-knockout mouse model with clear anatomical phenotype, single lab","pmids":["21584905"],"is_preprint":false},{"year":2007,"finding":"IL-1β induction of ADAMTS4 expression in human OA chondrocytes requires PKCζ (an atypical PKC); PKCζ mediates IL-1β-induced NF-κB activation (phosphorylation of IKKαβ and IκBα); pharmacological inhibition or siRNA/shRNA knockdown of PKCζ suppressed IL-1β-induced ADAMTS4 mRNA upregulation and aggrecanase activity.","method":"Pharmacological inhibitors of atypical PKCs; siRNA and shRNA knockdown of PKCζ; Western blotting for NF-κB pathway phosphorylation; quantitative PCR for ADAMTS4 mRNA; aggrecanase activity assay","journal":"Arthritis and rheumatism","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knockdown plus pharmacological inhibition with pathway readout and functional assay, single lab","pmids":["18050214"],"is_preprint":false},{"year":2009,"finding":"IL-1β induction of ADAMTS4 in chondrocytes requires MyD88, IRAK1, and TRAF6 adaptor proteins (each individually required but partial); Ras-mediated ROS production synergizes with these adaptors; combined knockdown of Ras and individual adaptors strongly blocked NF-κB activation (IKKαβ, IκBα phosphorylation) and ADAMTS4 induction.","method":"siRNA-mediated knockdown of MyD88, IRAK1, TRAF6, and Ras; antioxidant treatment; Western blotting for NF-κB pathway; ADAMTS4 mRNA quantitation","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — systematic siRNA knockdown with mechanistic pathway readout, single lab","pmids":["19342688"],"is_preprint":false},{"year":2016,"finding":"ADAMTS4 exerts anti-inflammatory effects in microglia and astrocytes; recombinant ADAMTS4 pretreatment of primary microglia and astrocytes decreased LPS-induced NO production and pro-inflammatory cytokine synthesis/release (NOS2, CCL2, TNF-α, IL-1β, MMP-9); siRNA silencing of ADAMTS4 increased these pro-inflammatory markers; in vivo, ADAMTS4 treatment decreased astrogliosis, macrophage infiltration, and increased M2 microglia after middle cerebral artery occlusion.","method":"Recombinant ADAMTS4 treatment of primary microglia/astrocyte cultures; siRNA silencing of ADAMTS4; NO and cytokine measurement; mouse middle cerebral artery occlusion model with ADAMTS4 treatment; immunohistochemistry","journal":"Glia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain- and loss-of-function in vitro with in vivo validation, single lab","pmids":["27301579"],"is_preprint":false},{"year":2013,"finding":"ADAMTS4 contributes to aggrecan degradation in human osteoarthritic nucleus pulposus cells non-redundantly with ADAMTS5; silencing either enzyme individually reduced TNF-α-dependent aggrecan degradation.","method":"Lentiviral shRNA silencing of ADAMTS-4 or ADAMTS-5 in human NP cells; Western blotting; aggrecan degradation assay","journal":"The American journal of pathology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — lentiviral knockdown of individual genes with functional aggrecan degradation readout, single lab","pmids":["23602832"],"is_preprint":false}],"current_model":"ADAMTS4 is a secreted, multi-domain metalloprotease (containing catalytic, disintegrin, thrombospondin, cysteine-rich, and spacer domains) that is synthesized as a latent zymogen, activated by furin/proprotein convertases in the trans-Golgi network and by GPI-anchored MT4-MMP-mediated C-terminal truncation on the cell surface; the activated enzyme cleaves aggrecan at multiple sites (predominantly in the chondroitin sulfate-rich region via exosite-mediated GAG binding through its TSP-1 motif and C-terminal domains), as well as versican, brevican, neurocan, hevin, Reelin, fibulin-2, and APP/Aβ; its activity is regulated by TIMP-3 (potent inhibitor), fibronectin (C-terminal domain inhibitor), CCN1 (cysteine-rich domain inhibitor), LRP1-mediated endocytosis, and transcriptionally by NF-κB, MAPK pathways, furin convertases, Runx2, NFATp, Sox4, and miR-125b; C-terminal spacer domain deletion releases the enzyme from the pericellular matrix and broadens its substrate specificity, while syndecan-1 anchors the activated p53 form to the cell surface."},"narrative":{"mechanistic_narrative":"ADAMTS4 (aggrecanase-1) is a secreted, multidomain metalloprotease that remodels the extracellular and pericellular matrix by cleaving large chondroitin sulfate proteoglycans, most prominently aggrecan, which it cuts at the Glu373-Ala374 interglobular-domain bond and at multiple more efficiently processed sites in the chondroitin sulfate-rich region [PMID:10751421, PMID:11854269]. It also cleaves versican (V1 at Glu441-Ala442 and many additional sites, V2 to generate brain GHAP), brevican, and other CNS proteoglycans, establishing distinct roles in connective tissue and neural matrix turnover [PMID:10986281, PMID:11278559, PMID:14561220, PMID:22420304, PMID:34450332]. Substrate recognition and activity depend critically on its C-terminal ancillary domains: the thrombospondin type-1 motif and cysteine-rich/spacer domains bind glycosaminoglycans to position substrate, sequential inclusion of these domains progressively enhances proteolysis, and aggrecan cleavage proceeds through an exosite mechanism in which substrate first docks at an exosite before the scissile peptide enters the active site [PMID:10827174, PMID:12202483, PMID:17430884, PMID:17487981]. The enzyme is produced as a latent zymogen activated by furin/proprotein convertase removal of the prodomain in the trans-Golgi and by GPI-anchored MT4-MMP (MMP-17)-mediated C-terminal truncation at the cell surface, where the active form is retained via syndecan-1; this activation step, rather than new synthesis, drives IL-1-induced cartilage aggrecanolysis [PMID:11796708, PMID:14744861, PMID:14701864, PMID:15780640]. The spacer domain confines full-length enzyme to the pericellular matrix and masks broad proteolytic activity, so its removal releases the enzyme and broadens substrate specificity [PMID:14662755, PMID:15161923]. ADAMTS4 activity is restrained by TIMP-3 (a potent inhibitor whose efficacy is enhanced by aggrecan GAGs), fibronectin (via the spacer domain), and CCN1 (via the cysteine-rich domain), and the enzyme is cleared by LRP1-mediated endocytosis [PMID:11278243, PMID:15161923, PMID:17470431, PMID:24474687, PMID:25709087]. Transcriptionally it is driven by inflammatory cytokine signaling through PKCζ, MyD88/IRAK1/TRAF6, MAPK and NF-κB pathways and by Runx2, NFATp, and Sox4, and is repressed post-transcriptionally by miR-125b [PMID:23602832, PMID:23406982, PMID:30016600, PMID:18050214, PMID:19342688]. Beyond matrix remodeling, ADAMTS4 cleaves APP/Aβ to generate N-truncated Aβ4-x peptides in oligodendrocytes, translocates to the nucleus of stressed smooth muscle cells to degrade PARP-1 and promote apoptosis in aortic aneurysm, and modulates neuroinflammation and CNS injury responses [PMID:28955046, PMID:30426203, PMID:27301579].","teleology":[{"year":2000,"claim":"Established the defining enzymatic identity of ADAMTS4 as aggrecanase-1 by mapping the precise aggrecan bonds it cleaves, distinguishing aggrecanase from MMP-type cleavage.","evidence":"In vitro cleavage of native aggrecan with recombinant ADAMTS4 and N-terminal sequencing of fragments","pmids":["10751421"],"confidence":"High","gaps":["Did not address how non-catalytic domains contribute to site selection","Cleavage efficiency in cell/tissue context not yet defined"]},{"year":2000,"claim":"Showed that substrate recognition is not purely active-site-driven but requires GAG-mediated binding through the thrombospondin type-1 motif, defining an ancillary-domain recognition mechanism.","evidence":"Domain truncation mutagenesis, TSP-1 peptide competition, and assays with GAG-free aggrecan","pmids":["10827174"],"confidence":"High","gaps":["Contributions of cysteine-rich and spacer domains to GAG binding not yet resolved","Structural basis of exosite binding not defined"]},{"year":2000,"claim":"Extended the substrate range beyond aggrecan to brevican, showing ADAMTS4 cleaves CNS proteoglycans at sites distinct from MMPs.","evidence":"In vitro digestion of brevican with purified ADAMTS4 and MMPs with N-terminal sequencing","pmids":["10986281"],"confidence":"High","gaps":["In vivo relevance of brevican cleavage not established","Cell type producing the enzyme in brain not identified"]},{"year":2001,"claim":"Identified TIMP-3 as the selective endogenous inhibitor of ADAMTS4, localizing the inhibitory activity to the TIMP-3 N-terminal domain.","evidence":"In vitro inhibition kinetics with recombinant TIMP domains","pmids":["11278243"],"confidence":"High","gaps":["How TIMP-3 engages the catalytic site structurally not resolved","Physiological regulation of TIMP-3/ADAMTS4 balance in tissue untested"]},{"year":2001,"claim":"Demonstrated versican as a physiological substrate and detected mature ADAMTS4 in vascular tissue, broadening its role to connective tissue beyond cartilage.","evidence":"In vitro versican cleavage with neoepitope antisera plus Western analysis of aortic intima","pmids":["11278559"],"confidence":"High","gaps":["Functional consequence of vascular versican cleavage not tested at this stage","Activation state of the detected tissue enzyme unknown"]},{"year":2002,"claim":"Defined the activation requirement for ADAMTS4: only C-terminally truncated p60/p53 forms are proteolytically active, and truncation is mediated by a GPI-anchored MMP, separating zymogen maturation from full enzymatic competence.","evidence":"Stable expression in chondrosarcoma cells, form fractionation with activity assays, and pharmacological inhibition","pmids":["11796708"],"confidence":"High","gaps":["Identity of the GPI-anchored MMP not yet established","Relationship of truncation to substrate specificity not defined"]},{"year":2002,"claim":"Established that C-terminal ancillary domains carry GAG-binding sites and that autocatalytic truncation reduces GAG affinity, linking domain composition to matrix anchoring.","evidence":"C-terminal sequencing of autocatalytic fragments and GAG-binding competition with synthetic peptides","pmids":["12202483"],"confidence":"High","gaps":["Physiological trigger for autocatalysis not defined","Quantitative impact on substrate cleavage in tissue not measured"]},{"year":2002,"claim":"Showed ADAMTS4 also cleaves the aggrecan MMP-site secondarily, and that this is TIMP-3-sensitive, reinforcing its mechanistic distinction from MMPs.","evidence":"In vitro digestion of aggrecan and G1-G2 constructs with selective TIMP inhibitors","pmids":["11854269"],"confidence":"High","gaps":["Biological significance of the secondary MMP-site cleavage unclear","Order/kinetics relative to primary cleavage not resolved in vivo"]},{"year":2003,"claim":"Revealed that the spacer domain masks general proteolytic activity and confines the enzyme to the pericellular matrix, so domain removal expands substrate range and releases the enzyme.","evidence":"Domain deletion mutants assayed against multiple substrates with localization analysis","pmids":["14662755"],"confidence":"High","gaps":["In vivo prevalence of spacer-deleted forms unknown","Mechanism of substrate masking at structural level not resolved here"]},{"year":2003,"claim":"Identified furin as the trans-Golgi prodomain-processing protease for pro-ADAMTS4 while noting a parallel furin-independent route.","evidence":"Co-localization, reciprocal co-IP, furin RNAi, and inhibitor/brefeldin A treatment","pmids":["14744861"],"confidence":"High","gaps":["Identity of the furin-independent pathway not defined","Whether prodomain removal alone confers activity not fully separated from C-terminal events"]},{"year":2003,"claim":"Identified MT4-MMP (MMP-17) as the GPI-anchored protease that truncates ADAMTS4 to the active p53 form at the cell surface, where syndecan-1 retains it.","evidence":"Co-transfection with active/inactive MT4-MMP, PI-PLC and GAG lyase treatments, and anti-syndecan-1 co-IP","pmids":["14701864"],"confidence":"High","gaps":["Regulation of MT4-MMP availability in tissue not addressed","Whether other surface proteases substitute in vivo unknown"]},{"year":2004,"claim":"Defined fibronectin as a spacer-domain-dependent inhibitor and co-localization partner, adding a matrix-based brake on aggrecanase activity.","evidence":"Yeast two-hybrid, cross-linking, solid-phase binding, and inhibition assays with domain mapping","pmids":["15161923"],"confidence":"High","gaps":["In vivo relevance of fibronectin inhibition not tested","Competition with substrate GAG binding not quantified"]},{"year":2004,"claim":"Linked ADAMTS4 to generation of the brain proteoglycan fragment GHAP via versican V2 cleavage, reinforcing a CNS matrix role.","evidence":"Anti-neoepitope analysis of purified GHAP and in vitro digestion of cerebellar proteoglycans","pmids":["14561220"],"confidence":"Medium","gaps":["In vivo enzyme responsible not genetically confirmed","Functional consequence of GHAP generation untested"]},{"year":2005,"claim":"Clarified the biochemical constraints on activation, showing prodomain removal under physiological conditions requires proprotein convertases (furin, PACE4, PC5/6) rather than autocatalysis.","evidence":"In vitro incubation of pro-ADAMTS4 with multiple proteases across conditions with activity assays","pmids":["16289022"],"confidence":"High","gaps":["Relative in vivo contribution of each convertase not defined","Integration with C-terminal MT4-MMP step not resolved"]},{"year":2005,"claim":"Demonstrated in cartilage explants that IL-1-driven aggrecanolysis depends on MT4-MMP-mediated ADAMTS4 activation rather than increased enzyme synthesis.","evidence":"Western analysis of ADAMTS4 species and aggrecan fragments with GPI/MMP pharmacological inhibition in explants","pmids":["15780640"],"confidence":"Medium","gaps":["Single-lab explant system; genetic confirmation absent","Distinction from ADAMTS5 contribution incomplete"]},{"year":2006,"claim":"Mapped the transcriptional control of the ADAMTS4 promoter in chondrocytes, identifying NFATp, Runx2 and a chondrocyte-specific NFI repressor element.","evidence":"Promoter-luciferase deletion reporter assays and mRNA analysis in chondrocytes","pmids":["16677612","11254106"],"confidence":"Medium","gaps":["Direct factor binding (e.g., ChIP) not shown for all sites","Cytokine-responsive elements not yet integrated"]},{"year":2007,"claim":"Systematized the domain hierarchy controlling activity and localization, showing sequential ancillary domains progressively enhance proteolysis and that the spacer domain governs ECM localization (versus cysteine-rich for ADAMTS5).","evidence":"Comparative domain-deletion mutagenesis with multi-substrate assays against ADAMTS5","pmids":["17430884"],"confidence":"High","gaps":["Structural mechanism of domain cooperation not resolved here","In vivo consequences of domain configuration untested"]},{"year":2007,"claim":"Provided structural and kinetic mechanisms: crystal structures revealed open/closed catalytic configurations, and kinetics defined an exosite-mediated aggrecan cleavage with implications for inhibitor design.","evidence":"X-ray crystallography of apo/inhibitor-bound enzyme; enzyme kinetics with aggrecan and peptide substrates plus active-site inhibitor","pmids":["18042673","17487981"],"confidence":"High","gaps":["Substrate-bound structure not solved","Physiological trigger switching open/closed states unknown"]},{"year":2007,"claim":"Showed aggrecan GAG binding enhances TIMP-3 inhibition, revealing substrate-inhibitor interplay at the ancillary domains.","evidence":"FRET peptide assay with aggrecan-binding competition and solid-phase binding","pmids":["17470431"],"confidence":"Medium","gaps":["Single-lab mechanistic study; in vivo relevance untested","Structural basis of the ternary complex unresolved"]},{"year":2009,"claim":"Identified upstream cytokine signaling requirements (PKCζ, MyD88/IRAK1/TRAF6, Ras/ROS) for IL-1β-driven ADAMTS4 induction via NF-κB.","evidence":"siRNA/shRNA knockdown and pharmacological inhibition with NF-κB pathway and ADAMTS4 mRNA readouts in chondrocytes","pmids":["18050214","19342688"],"confidence":"Medium","gaps":["Promoter elements engaged by these pathways not fully mapped","Single-lab studies; in vivo validation limited"]},{"year":2009,"claim":"Extended substrate range to the matricellular protein hevin and linked ADAMTS4 proteolysis to cerebellar development in vivo.","evidence":"In vitro hevin digestion and in vivo co-localization in mouse cerebellum","pmids":["20018883"],"confidence":"Medium","gaps":["Genetic loss-of-function effect on hevin in vivo not shown","Functional outcome of the SPARC-like fragment unresolved"]},{"year":2011,"claim":"Revealed a developmental role through genetic redundancy: Adamts4 alone is dispensable, but Adamts1/Adamts4 double knockouts die perinatally with renal medulla defects.","evidence":"Gene targeting and genetic cross with histological renal analysis","pmids":["21584905"],"confidence":"Medium","gaps":["Molecular substrate underlying the renal phenotype unidentified","Extent of redundancy with other ADAMTS members untested"]},{"year":2012,"claim":"Established LRP1-mediated endocytosis as the clearance route for ADAMTS4 via cysteine-rich/spacer binding, with slower turnover than ADAMTS5.","evidence":"Domain mutagenesis, soluble LRP1 cluster binding affinity, and endocytosis kinetics with competition assays","pmids":["24474687"],"confidence":"High","gaps":["In vivo contribution of LRP1 clearance to matrix turnover not quantified","Regulation of LRP1 availability not addressed"]},{"year":2012,"claim":"Demonstrated CNS therapeutic relevance: ADAMTS4 degrades inhibitory CSPGs (brevican, neurocan, phosphacan) and promotes axonal regeneration and motor recovery after spinal cord injury.","evidence":"In vitro CSPG degradation and neurite outgrowth assays plus in vivo spinal cord contusion model","pmids":["22420304"],"confidence":"Medium","gaps":["Endogenous versus exogenous enzyme contribution not separated","Long-term and mechanistic basis of recovery not fully defined"]},{"year":2012,"claim":"Added Reelin to the substrate repertoire under multi-inhibitor control, connecting ADAMTS4 to hippocampal Reelin processing.","evidence":"In vitro cleavage with inhibitor dissection and hippocampal immunolocalization","pmids":["23082219"],"confidence":"Medium","gaps":["In vivo Reelin processing by ADAMTS4 genetically unconfirmed","Functional consequence for Reelin signaling untested"]},{"year":2013,"claim":"Defined cytokine-driven transcriptional control in nucleus pulposus cells through MAPK/NF-κB and showed ADAMTS4 contributes non-redundantly to aggrecan degradation alongside ADAMTS5.","evidence":"Promoter assays, pathway inhibitors, lentiviral shRNA silencing, and aggrecan degradation assays","pmids":["23602832","23406982"],"confidence":"Medium","gaps":["miR-125b regulation shown by reporter, not in vivo","Relative ADAMTS4/ADAMTS5 contribution context-dependent"]},{"year":2015,"claim":"Identified CCN1 as a cysteine-rich-domain-binding inhibitor that keeps ADAMTS4 latent, revealing an additional endogenous brake unmasked by CCN1 knockdown.","evidence":"LC-MS/MS co-purification, immunoprecipitation, solid-phase binding, aggrecan digestion, and siRNA knockdown","pmids":["25709087"],"confidence":"Medium","gaps":["Structural basis of CCN1 inhibition unresolved","In vivo relevance to OA progression untested"]},{"year":2016,"claim":"Uncovered an anti-inflammatory role in CNS glia, where ADAMTS4 suppresses microglial/astrocyte pro-inflammatory responses and improves outcome after cerebral ischemia.","evidence":"Recombinant treatment and siRNA silencing in glia with cytokine measurement and a middle cerebral artery occlusion model","pmids":["27301579"],"confidence":"Medium","gaps":["Molecular mechanism of anti-inflammatory action undefined","Substrate mediating glial effects unidentified"]},{"year":2017,"claim":"Revealed a non-canonical intracellular function: stress-induced nuclear translocation and PARP-1 cleavage driving smooth muscle apoptosis in aortic aneurysm/dissection.","evidence":"Adamts4-/- mouse aneurysm model, nuclear translocation imaging, and direct PARP-1 cleavage assay","pmids":["28955046"],"confidence":"Medium","gaps":["Mechanism of nuclear import for a secreted protease unexplained","PARP-1 cleavage relies on a single study"]},{"year":2018,"claim":"Established ADAMTS4 as an oligodendrocyte-specific generator of N-truncated Aβ4-x peptides, implicating it in amyloid processing.","evidence":"Recognition-site mapping, inducible overexpression with Aβ ELISA, ADAMTS4-/- mice, and ADAMTS4-/- oligodendrocyte cultures","pmids":["30426203"],"confidence":"High","gaps":["Contribution to amyloid pathology in human disease not established","Regulation of oligodendrocyte ADAMTS4 expression unclear"]},{"year":2018,"claim":"Identified Sox4 (and Sox11) as direct transcriptional activators of ADAMTS4 promoting cartilage destruction.","evidence":"Luciferase reporter, ChIP, and adenoviral overexpression in femoral head organ cultures","pmids":["30016600"],"confidence":"Medium","gaps":["Integration with cytokine-driven regulation not mapped","In vivo Sox4-ADAMTS4 axis in OA untested"]},{"year":2020,"claim":"Linked ADAMTS4 to acute infection biology, showing damage-responsive lung fibroblasts produce it to remodel ECM and promote immune infiltration, with levels tracking influenza severity.","evidence":"Single-cell transcriptomics, mouse influenza model, and human BALF measurements across cohorts","pmids":["33116313"],"confidence":"Medium","gaps":["Specific ECM substrate driving immune infiltration not defined","Causal mechanism in humans remains correlative"]},{"year":2021,"claim":"Refined substrate-site preference by mapping 21 novel versican V1 cleavage sites and a P1-Glu preference, expanding the proteolytic landscape.","evidence":"In vitro versican digestion with active versus inactive enzyme and label-free LC-MS/MS proteomics","pmids":["34450332"],"confidence":"Medium","gaps":["Biological significance of individual novel sites untested","In vivo occurrence of these cleavages not confirmed"]},{"year":null,"claim":"How a secreted matrix protease achieves nuclear localization and intracellular PARP-1 cleavage, and how its diverse extracellular and intracellular activities are coordinated across tissues, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No mechanism for nuclear import of ADAMTS4 established","Integration of matrix-remodeling, anti-inflammatory, and apoptotic functions not unified","Substrate driving most in vivo phenotypes often unidentified"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,2,3,21,28,29,32]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0,5,13]},{"term_id":"GO:0140098","term_label":"catalytic activity, acting on RNA","supporting_discovery_ids":[1,6]}],"localization":[{"term_id":"GO:0005576","term_label":"extracellular region","supporting_discovery_ids":[3,5,31]},{"term_id":"GO:0031012","term_label":"extracellular matrix","supporting_discovery_ids":[8,11,15]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[10,5]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[9]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[28]}],"pathway":[{"term_id":"R-HSA-1474244","term_label":"Extracellular matrix organization","supporting_discovery_ids":[0,3,8]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[5,9,13]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[28,29,31]}],"complexes":[],"partners":["TIMP3","MMP17","SDC1","FN1","CCN1","LRP1","FURIN"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O75173","full_name":"A disintegrin and metalloproteinase with thrombospondin motifs 4","aliases":["ADMP-1","Aggrecanase-1"],"length_aa":837,"mass_kda":90.2,"function":"Cleaves aggrecan, a cartilage proteoglycan, at the '392-Glu-|-Ala-393' site and may be involved in its turnover (PubMed:10356395, PubMed:10827174). Also cleaves COMP (PubMed:39672391). May play an important role in the destruction of aggrecan in arthritic diseases. Could be a critical factor in the exacerbation of neurodegeneration in Alzheimer disease","subcellular_location":"Secreted, extracellular space, extracellular matrix","url":"https://www.uniprot.org/uniprotkb/O75173/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ADAMTS4","classification":"Not Classified","n_dependent_lines":10,"n_total_lines":1208,"dependency_fraction":0.008278145695364239},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/ADAMTS4","total_profiled":1310},"omim":[{"mim_id":"605421","title":"A DISINTEGRIN-LIKE AND METALLOPROTEINASE WITH THROMBOSPONDIN TYPE 1 MOTIF, 9; ADAMTS9","url":"https://www.omim.org/entry/605421"},{"mim_id":"605011","title":"A DISINTEGRIN-LIKE AND METALLOPROTEINASE WITH THROMBOSPONDIN TYPE 1 MOTIF, 3; ADAMTS3","url":"https://www.omim.org/entry/605011"},{"mim_id":"605009","title":"A DISINTEGRIN-LIKE AND METALLOPROTEINASE WITH THROMBOSPONDIN TYPE 1 MOTIF, 7; ADAMTS7","url":"https://www.omim.org/entry/605009"},{"mim_id":"605008","title":"A DISINTEGRIN-LIKE AND METALLOPROTEINASE WITH THROMBOSPONDIN TYPE 1 MOTIF, 6; ADAMTS6","url":"https://www.omim.org/entry/605008"},{"mim_id":"605007","title":"A DISINTEGRIN-LIKE AND METALLOPROTEINASE WITH THROMBOSPONDIN TYPE 1 MOTIF, 5; ADAMTS5","url":"https://www.omim.org/entry/605007"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nuclear speckles","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"adipose tissue","ntpm":108.5},{"tissue":"ovary","ntpm":84.8}],"url":"https://www.proteinatlas.org/search/ADAMTS4"},"hgnc":{"alias_symbol":["KIAA0688","ADAMTS-2","ADMP-1"],"prev_symbol":[]},"alphafold":{"accession":"O75173","domains":[{"cath_id":"3.40.390.10","chopping":"215-419","consensus_level":"high","plddt":89.4455,"start":215,"end":419},{"cath_id":"3.40.1620.60","chopping":"440-516","consensus_level":"high","plddt":89.736,"start":440,"end":516},{"cath_id":"-","chopping":"583-640","consensus_level":"medium","plddt":85.5107,"start":583,"end":640},{"cath_id":"2.60.120.830","chopping":"686-804","consensus_level":"high","plddt":90.7829,"start":686,"end":804},{"cath_id":"2.40.128","chopping":"60-166_177-183","consensus_level":"high","plddt":67.7167,"start":60,"end":183}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O75173","model_url":"https://alphafold.ebi.ac.uk/files/AF-O75173-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O75173-F1-predicted_aligned_error_v6.png","plddt_mean":80.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ADAMTS4","jax_strain_url":"https://www.jax.org/strain/search?query=ADAMTS4"},"sequence":{"accession":"O75173","fasta_url":"https://rest.uniprot.org/uniprotkb/O75173.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O75173/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O75173"}},"corpus_meta":[{"pmid":"11278243","id":"PMC_11278243","title":"TIMP-3 is a potent inhibitor of aggrecanase 1 (ADAM-TS4) and aggrecanase 2 (ADAM-TS5).","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11278243","citation_count":404,"is_preprint":false},{"pmid":"11278559","id":"PMC_11278559","title":"Versican V1 proteolysis in human aorta in vivo occurs at the Glu441-Ala442 bond, a site that is cleaved by recombinant ADAMTS-1 and ADAMTS-4.","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11278559","citation_count":389,"is_preprint":false},{"pmid":"17265492","id":"PMC_17265492","title":"Aggrecan degradation in human articular cartilage explants is mediated by both ADAMTS-4 and ADAMTS-5.","date":"2007","source":"Arthritis and rheumatism","url":"https://pubmed.ncbi.nlm.nih.gov/17265492","citation_count":334,"is_preprint":false},{"pmid":"12568957","id":"PMC_12568957","title":"IL-1 beta induces COX2, MMP-1, -3 and -13, ADAMTS-4, IL-1 beta and IL-6 in human tendon cells.","date":"2003","source":"Journal of orthopaedic research : official publication of the Orthopaedic Research Society","url":"https://pubmed.ncbi.nlm.nih.gov/12568957","citation_count":300,"is_preprint":false},{"pmid":"11956193","id":"PMC_11956193","title":"Inhibition of ADAM-TS4 and ADAM-TS5 prevents aggrecan degradation in osteoarthritic cartilage.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11956193","citation_count":241,"is_preprint":false},{"pmid":"17430884","id":"PMC_17430884","title":"Proteolytic activities of human ADAMTS-5: comparative studies with ADAMTS-4.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17430884","citation_count":219,"is_preprint":false},{"pmid":"10751421","id":"PMC_10751421","title":"Sites of aggrecan cleavage by recombinant human aggrecanase-1 (ADAMTS-4).","date":"2000","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10751421","citation_count":197,"is_preprint":false},{"pmid":"33116313","id":"PMC_33116313","title":"Exuberant fibroblast activity compromises lung function via ADAMTS4.","date":"2020","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/33116313","citation_count":176,"is_preprint":false},{"pmid":"14662755","id":"PMC_14662755","title":"Altered proteolytic activities of ADAMTS-4 expressed by C-terminal processing.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/14662755","citation_count":176,"is_preprint":false},{"pmid":"23602832","id":"PMC_23602832","title":"Inflammatory cytokines associated with degenerative disc disease control aggrecanase-1 (ADAMTS-4) expression in nucleus pulposus cells through MAPK and NF-κB.","date":"2013","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/23602832","citation_count":166,"is_preprint":false},{"pmid":"10827174","id":"PMC_10827174","title":"The thrombospondin motif of aggrecanase-1 (ADAMTS-4) is critical for aggrecan substrate recognition and cleavage.","date":"2000","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10827174","citation_count":163,"is_preprint":false},{"pmid":"14701864","id":"PMC_14701864","title":"ADAMTS4 (aggrecanase-1) activation on the cell surface involves C-terminal cleavage by glycosylphosphatidyl inositol-anchored membrane type 4-matrix metalloproteinase and binding of the activated proteinase to chondroitin sulfate and heparan sulfate on syndecan-1.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/14701864","citation_count":156,"is_preprint":false},{"pmid":"18328163","id":"PMC_18328163","title":"The regulation of the ADAMTS4 and ADAMTS5 aggrecanases in osteoarthritis: a review.","date":"2008","source":"Clinical and experimental rheumatology","url":"https://pubmed.ncbi.nlm.nih.gov/18328163","citation_count":155,"is_preprint":false},{"pmid":"11741898","id":"PMC_11741898","title":"Cloning and characterization of ADAMTS-14, a novel ADAMTS displaying high homology with ADAMTS-2 and ADAMTS-3.","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11741898","citation_count":154,"is_preprint":false},{"pmid":"11796708","id":"PMC_11796708","title":"Activation of the proteolytic activity of ADAMTS4 (aggrecanase-1) by C-terminal truncation.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11796708","citation_count":153,"is_preprint":false},{"pmid":"10986281","id":"PMC_10986281","title":"Brevican is degraded by matrix metalloproteinases and aggrecanase-1 (ADAMTS4) at different sites.","date":"2000","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10986281","citation_count":140,"is_preprint":false},{"pmid":"12202483","id":"PMC_12202483","title":"Autocatalytic cleavage of ADAMTS-4 (Aggrecanase-1) reveals multiple glycosaminoglycan-binding sites.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12202483","citation_count":123,"is_preprint":false},{"pmid":"12392761","id":"PMC_12392761","title":"Characterization of human aggrecanase 2 (ADAM-TS5): substrate specificity studies and comparison with aggrecanase 1 (ADAM-TS4).","date":"2002","source":"Matrix biology : journal of the International Society for Matrix Biology","url":"https://pubmed.ncbi.nlm.nih.gov/12392761","citation_count":120,"is_preprint":false},{"pmid":"18662930","id":"PMC_18662930","title":"Hyaluronan inhibits expression of ADAMTS4 (aggrecanase-1) in human osteoarthritic chondrocytes.","date":"2008","source":"Annals of the rheumatic diseases","url":"https://pubmed.ncbi.nlm.nih.gov/18662930","citation_count":113,"is_preprint":false},{"pmid":"18042673","id":"PMC_18042673","title":"Crystal structures of the two major aggrecan degrading enzymes, ADAMTS4 and ADAMTS5.","date":"2007","source":"Protein science : a publication of the Protein Society","url":"https://pubmed.ncbi.nlm.nih.gov/18042673","citation_count":111,"is_preprint":false},{"pmid":"16003758","id":"PMC_16003758","title":"Matrix-degrading proteases ADAMTS4 and ADAMTS5 (disintegrins and metalloproteinases with thrombospondin motifs 4 and 5) are expressed in human glioblastomas.","date":"2006","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/16003758","citation_count":106,"is_preprint":false},{"pmid":"17606262","id":"PMC_17606262","title":"ADAMTS-4 and -8 are inflammatory regulated enzymes expressed in macrophage-rich areas of human atherosclerotic plaques.","date":"2007","source":"Atherosclerosis","url":"https://pubmed.ncbi.nlm.nih.gov/17606262","citation_count":100,"is_preprint":false},{"pmid":"17922681","id":"PMC_17922681","title":"Expression of ADAMTS4 (aggrecanase-1) in human osteoarthritic cartilage.","date":"2007","source":"Pathology international","url":"https://pubmed.ncbi.nlm.nih.gov/17922681","citation_count":98,"is_preprint":false},{"pmid":"14744861","id":"PMC_14744861","title":"Proprotein convertase furin interacts with and cleaves pro-ADAMTS4 (Aggrecanase-1) in the trans-Golgi network.","date":"2004","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/14744861","citation_count":98,"is_preprint":false},{"pmid":"11284712","id":"PMC_11284712","title":"Transgenic mice with inactive alleles for procollagen N-proteinase (ADAMTS-2) develop fragile skin and male sterility.","date":"2001","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/11284712","citation_count":94,"is_preprint":false},{"pmid":"18941754","id":"PMC_18941754","title":"Expression of ADAMTS-4 by chondrocytes in the surface zone of human osteoarthritic cartilage is regulated by epigenetic DNA de-methylation.","date":"2008","source":"Rheumatology international","url":"https://pubmed.ncbi.nlm.nih.gov/18941754","citation_count":90,"is_preprint":false},{"pmid":"12646579","id":"PMC_12646579","title":"Transforming growth factor-beta induces secretion of activated ADAMTS-2. A procollagen III N-proteinase.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12646579","citation_count":90,"is_preprint":false},{"pmid":"16046392","id":"PMC_16046392","title":"Domains and maturation processes that regulate the activity of ADAMTS-2, a metalloproteinase cleaving the aminopropeptide of fibrillar procollagens types I-III and V.","date":"2005","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16046392","citation_count":90,"is_preprint":false},{"pmid":"23406982","id":"PMC_23406982","title":"MicroRNA-125b regulates the expression of aggrecanase-1 (ADAMTS-4) in human osteoarthritic chondrocytes.","date":"2013","source":"Arthritis research & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/23406982","citation_count":89,"is_preprint":false},{"pmid":"12528112","id":"PMC_12528112","title":"Induction of aggrecanase 1 (ADAM-TS4) by interleukin-1 occurs through activation of constitutively produced protein.","date":"2003","source":"Arthritis and rheumatism","url":"https://pubmed.ncbi.nlm.nih.gov/12528112","citation_count":82,"is_preprint":false},{"pmid":"14561220","id":"PMC_14561220","title":"ADAMTS4 (aggrecanase-1) cleaves human brain versican V2 at Glu405-Gln406 to generate glial hyaluronate binding protein.","date":"2004","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/14561220","citation_count":79,"is_preprint":false},{"pmid":"22420304","id":"PMC_22420304","title":"The endogenous proteoglycan-degrading enzyme ADAMTS-4 promotes functional recovery after spinal cord injury.","date":"2012","source":"Journal of neuroinflammation","url":"https://pubmed.ncbi.nlm.nih.gov/22420304","citation_count":77,"is_preprint":false},{"pmid":"24474687","id":"PMC_24474687","title":"Low density lipoprotein receptor-related protein 1 (LRP1)-mediated endocytic clearance of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4): functional differences of non-catalytic domains of ADAMTS-4 and ADAMTS-5 in LRP1 binding.","date":"2014","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24474687","citation_count":76,"is_preprint":false},{"pmid":"23245439","id":"PMC_23245439","title":"ADAMTS-1 and ADAMTS-4 levels are elevated in thoracic aortic aneurysms and dissections.","date":"2012","source":"The Annals of thoracic surgery","url":"https://pubmed.ncbi.nlm.nih.gov/23245439","citation_count":72,"is_preprint":false},{"pmid":"15780640","id":"PMC_15780640","title":"Analysis of ADAMTS4 and MT4-MMP indicates that both are involved in aggrecanolysis in interleukin-1-treated bovine cartilage.","date":"2005","source":"Osteoarthritis and cartilage","url":"https://pubmed.ncbi.nlm.nih.gov/15780640","citation_count":70,"is_preprint":false},{"pmid":"28955046","id":"PMC_28955046","title":"Critical Role of ADAMTS-4 in the Development of Sporadic Aortic Aneurysm and Dissection in Mice.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28955046","citation_count":70,"is_preprint":false},{"pmid":"16771712","id":"PMC_16771712","title":"TIMP-3 inhibits the procollagen N-proteinase ADAMTS-2.","date":"2006","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/16771712","citation_count":70,"is_preprint":false},{"pmid":"23082219","id":"PMC_23082219","title":"Regulated proteolytic processing of Reelin through interplay of tissue plasminogen activator (tPA), ADAMTS-4, ADAMTS-5, and their modulators.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23082219","citation_count":69,"is_preprint":false},{"pmid":"11854269","id":"PMC_11854269","title":"ADAMTS4 cleaves at the aggrecanase site (Glu373-Ala374) and secondarily at the matrix metalloproteinase site (Asn341-Phe342) in the aggrecan interglobular domain.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11854269","citation_count":68,"is_preprint":false},{"pmid":"25045124","id":"PMC_25045124","title":"Leptin induces ADAMTS-4, ADAMTS-5, and ADAMTS-9 genes expression by mitogen-activated protein kinases and NF-ĸB signaling pathways in human chondrocytes.","date":"2014","source":"Cell biology international","url":"https://pubmed.ncbi.nlm.nih.gov/25045124","citation_count":65,"is_preprint":false},{"pmid":"26288689","id":"PMC_26288689","title":"Discovery of Potent and Selective Inhibitors for ADAMTS-4 through DNA-Encoded Library Technology (ELT).","date":"2015","source":"ACS medicinal chemistry letters","url":"https://pubmed.ncbi.nlm.nih.gov/26288689","citation_count":64,"is_preprint":false},{"pmid":"15161923","id":"PMC_15161923","title":"ADAMTS4 (aggrecanase-1) interaction with the C-terminal domain of fibronectin inhibits proteolysis of aggrecan.","date":"2004","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/15161923","citation_count":59,"is_preprint":false},{"pmid":"20574651","id":"PMC_20574651","title":"ADAMTS-2 functions as anti-angiogenic and anti-tumoral molecule independently of its catalytic activity.","date":"2010","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/20574651","citation_count":59,"is_preprint":false},{"pmid":"24126638","id":"PMC_24126638","title":"Tumor necrosis factor-α induces ADAMTS-4 expression in human osteoarthritis chondrocytes.","date":"2013","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/24126638","citation_count":59,"is_preprint":false},{"pmid":"16289022","id":"PMC_16289022","title":"ADAMTS-4 (aggrecanase-1): N-terminal activation mechanisms.","date":"2005","source":"Archives of biochemistry and biophysics","url":"https://pubmed.ncbi.nlm.nih.gov/16289022","citation_count":57,"is_preprint":false},{"pmid":"17470431","id":"PMC_17470431","title":"TIMP-3 inhibition of ADAMTS-4 (Aggrecanase-1) is modulated by interactions between aggrecan and the C-terminal domain of ADAMTS-4.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17470431","citation_count":57,"is_preprint":false},{"pmid":"20645923","id":"PMC_20645923","title":"Reactive-site mutants of N-TIMP-3 that selectively inhibit ADAMTS-4 and ADAMTS-5: biological and structural implications.","date":"2010","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/20645923","citation_count":56,"is_preprint":false},{"pmid":"16677612","id":"PMC_16677612","title":"Regulation of the human ADAMTS-4 promoter by transcription factors and cytokines.","date":"2006","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/16677612","citation_count":52,"is_preprint":false},{"pmid":"27491335","id":"PMC_27491335","title":"Loss of ADAMTS4 reduces high fat diet-induced atherosclerosis and enhances plaque stability in ApoE(-/-) mice.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/27491335","citation_count":51,"is_preprint":false},{"pmid":"30426203","id":"PMC_30426203","title":"The metalloprotease ADAMTS4 generates N-truncated Aβ4-x species and marks oligodendrocytes as a source of amyloidogenic peptides in Alzheimer's disease.","date":"2018","source":"Acta neuropathologica","url":"https://pubmed.ncbi.nlm.nih.gov/30426203","citation_count":51,"is_preprint":false},{"pmid":"30016600","id":"PMC_30016600","title":"Sox4 is involved in osteoarthritic cartilage deterioration through induction of ADAMTS4 and ADAMTS5.","date":"2018","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/30016600","citation_count":49,"is_preprint":false},{"pmid":"18387286","id":"PMC_18387286","title":"The regulation of aggrecanase ADAMTS-4 expression in human Achilles tendon and tendon-derived cells.","date":"2008","source":"Matrix biology : journal of the International Society for Matrix Biology","url":"https://pubmed.ncbi.nlm.nih.gov/18387286","citation_count":49,"is_preprint":false},{"pmid":"15457452","id":"PMC_15457452","title":"Release of hyaluronan and hyaladherins (aggrecan G1 domain and link proteins) from articular cartilage exposed to ADAMTS-4 (aggrecanase 1) or ADAMTS-5 (aggrecanase 2).","date":"2004","source":"Arthritis and rheumatism","url":"https://pubmed.ncbi.nlm.nih.gov/15457452","citation_count":43,"is_preprint":false},{"pmid":"24732836","id":"PMC_24732836","title":"Upregulation of tumor necrosis factor α and ADAMTS-5, but not ADAMTS-4, in human intervertebral cartilage endplate with modic changes.","date":"2014","source":"Spine","url":"https://pubmed.ncbi.nlm.nih.gov/24732836","citation_count":42,"is_preprint":false},{"pmid":"25501175","id":"PMC_25501175","title":"Increased serum ADAMTS-4 in knee osteoarthritis: a potential indicator for the diagnosis of osteoarthritis in early stages.","date":"2014","source":"Genetics and molecular research : GMR","url":"https://pubmed.ncbi.nlm.nih.gov/25501175","citation_count":41,"is_preprint":false},{"pmid":"20662062","id":"PMC_20662062","title":"Cytokine-induced increases in ADAMTS-4 messenger RNA expression do not lead to increased aggrecanase activity in ADAMTS-5-deficient mice.","date":"2010","source":"Arthritis and rheumatism","url":"https://pubmed.ncbi.nlm.nih.gov/20662062","citation_count":41,"is_preprint":false},{"pmid":"34450332","id":"PMC_34450332","title":"Identification of novel ADAMTS1, ADAMTS4 and ADAMTS5 cleavage sites in versican using a label-free quantitative proteomics approach.","date":"2021","source":"Journal of proteomics","url":"https://pubmed.ncbi.nlm.nih.gov/34450332","citation_count":38,"is_preprint":false},{"pmid":"23438438","id":"PMC_23438438","title":"Mechanisms involved in suppression of ADAMTS4 expression in synoviocytes by high molecular weight hyaluronic acid.","date":"2013","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/23438438","citation_count":38,"is_preprint":false},{"pmid":"27301579","id":"PMC_27301579","title":"Anti-inflammatory effects of ADAMTS-4 in a mouse model of ischemic stroke.","date":"2016","source":"Glia","url":"https://pubmed.ncbi.nlm.nih.gov/27301579","citation_count":38,"is_preprint":false},{"pmid":"10961658","id":"PMC_10961658","title":"ADAMTS-4 (a disintegrin and metalloproteinase with thrombospondin motifs) is transcriptionally induced in beta-amyloid treated rat astrocytes.","date":"2000","source":"Neuroscience letters","url":"https://pubmed.ncbi.nlm.nih.gov/10961658","citation_count":38,"is_preprint":false},{"pmid":"23319426","id":"PMC_23319426","title":"ADAMTS4 and its proteolytic fragments differentially affect melanoma growth and angiogenesis in mice.","date":"2013","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/23319426","citation_count":34,"is_preprint":false},{"pmid":"32044329","id":"PMC_32044329","title":"Astrocyte-selective AAV-ADAMTS4 gene therapy combined with hindlimb rehabilitation promotes functional recovery after spinal cord injury.","date":"2020","source":"Experimental neurology","url":"https://pubmed.ncbi.nlm.nih.gov/32044329","citation_count":34,"is_preprint":false},{"pmid":"16741450","id":"PMC_16741450","title":"Expression of ADAMTS-4 (aggrecanase-1) and possible involvement in regression of lumbar disc herniation.","date":"2006","source":"Spine","url":"https://pubmed.ncbi.nlm.nih.gov/16741450","citation_count":34,"is_preprint":false},{"pmid":"22264287","id":"PMC_22264287","title":"Anti-angiogenic properties of ADAMTS-4 in vitro.","date":"2012","source":"International journal of experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/22264287","citation_count":33,"is_preprint":false},{"pmid":"24595230","id":"PMC_24595230","title":"Pentosan polysulfate decreases myocardial expression of the extracellular matrix enzyme ADAMTS4 and improves cardiac function in vivo in rats subjected to pressure overload by aortic banding.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24595230","citation_count":33,"is_preprint":false},{"pmid":"25709087","id":"PMC_25709087","title":"CCN1 (Cyr61) Is Overexpressed in Human Osteoarthritic Cartilage and Inhibits ADAMTS-4 (Aggrecanase 1) Activity.","date":"2015","source":"Arthritis & rheumatology (Hoboken, N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/25709087","citation_count":33,"is_preprint":false},{"pmid":"22324945","id":"PMC_22324945","title":"Interleukin-6 upregulates expression of ADAMTS-4 in fibroblast-like synoviocytes from patients with rheumatoid arthritis.","date":"2011","source":"International journal of rheumatic diseases","url":"https://pubmed.ncbi.nlm.nih.gov/22324945","citation_count":32,"is_preprint":false},{"pmid":"26495885","id":"PMC_26495885","title":"Association between ADAMTS-4 gene polymorphism and lumbar disc degeneration in Chinese Han population.","date":"2015","source":"Journal of orthopaedic research : official publication of the Orthopaedic Research Society","url":"https://pubmed.ncbi.nlm.nih.gov/26495885","citation_count":31,"is_preprint":false},{"pmid":"19506088","id":"PMC_19506088","title":"Cell death-associated ADAMTS4 and versican degradation in vascular tissue.","date":"2009","source":"The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society","url":"https://pubmed.ncbi.nlm.nih.gov/19506088","citation_count":29,"is_preprint":false},{"pmid":"20018883","id":"PMC_20018883","title":"Processing of the matricellular protein hevin in mouse brain is dependent on ADAMTS4.","date":"2009","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20018883","citation_count":29,"is_preprint":false},{"pmid":"17487981","id":"PMC_17487981","title":"Substrate-dependent inhibition kinetics of an active site-directed inhibitor of ADAMTS-4 (Aggrecanase 1).","date":"2007","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17487981","citation_count":29,"is_preprint":false},{"pmid":"32469162","id":"PMC_32469162","title":"MiR-126a-5p limits the formation of abdominal aortic aneurysm in mice and decreases ADAMTS-4 expression.","date":"2020","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/32469162","citation_count":28,"is_preprint":false},{"pmid":"28099917","id":"PMC_28099917","title":"Cleavage of Fibulin-2 by the aggrecanases ADAMTS-4 and ADAMTS-5 contributes to the tumorigenic potential of breast cancer cells.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/28099917","citation_count":28,"is_preprint":false},{"pmid":"18050214","id":"PMC_18050214","title":"Involvement of protein kinase Czeta in interleukin-1beta induction of ADAMTS-4 and type 2 nitric oxide synthase via NF-kappaB signaling in primary human osteoarthritic chondrocytes.","date":"2007","source":"Arthritis and rheumatism","url":"https://pubmed.ncbi.nlm.nih.gov/18050214","citation_count":28,"is_preprint":false},{"pmid":"23778376","id":"PMC_23778376","title":"Assessment of the matrix degenerative effects of MMP-3, ADAMTS-4, and HTRA1, injected into a bovine intervertebral disc organ culture model.","date":"2013","source":"Spine","url":"https://pubmed.ncbi.nlm.nih.gov/23778376","citation_count":28,"is_preprint":false},{"pmid":"26809777","id":"PMC_26809777","title":"ADAMTS-4 promotes neurodegeneration in a mouse model of amyotrophic lateral sclerosis.","date":"2016","source":"Molecular neurodegeneration","url":"https://pubmed.ncbi.nlm.nih.gov/26809777","citation_count":27,"is_preprint":false},{"pmid":"20857147","id":"PMC_20857147","title":"Transcript levels of major MMPs and ADAMTS-4 in relation to the clinicopathological profile of patients with lumbar disc herniation.","date":"2010","source":"European spine journal : official publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society","url":"https://pubmed.ncbi.nlm.nih.gov/20857147","citation_count":27,"is_preprint":false},{"pmid":"29694979","id":"PMC_29694979","title":"Promotion of Tumor Growth by ADAMTS4 in Colorectal Cancer: Focused on Macrophages.","date":"2018","source":"Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/29694979","citation_count":26,"is_preprint":false},{"pmid":"24752352","id":"PMC_24752352","title":"IL-6 upregulates a disintegrin and metalloproteinase with thrombospondin motifs 2 (ADAMTS-2) in human osteosarcoma cells mediated by JNK pathway.","date":"2014","source":"Molecular and cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24752352","citation_count":25,"is_preprint":false},{"pmid":"15262240","id":"PMC_15262240","title":"Effect of adenovirus-mediated overexpression of bovine ADAMTS-4 and human ADAMTS-5 in primary bovine articular chondrocyte pellet culture system.","date":"2004","source":"Osteoarthritis and cartilage","url":"https://pubmed.ncbi.nlm.nih.gov/15262240","citation_count":25,"is_preprint":false},{"pmid":"29153440","id":"PMC_29153440","title":"Relationship between ADAMTS4 and carotid atherosclerotic plaque vulnerability in humans.","date":"2017","source":"Journal of vascular surgery","url":"https://pubmed.ncbi.nlm.nih.gov/29153440","citation_count":24,"is_preprint":false},{"pmid":"33350314","id":"PMC_33350314","title":"Interrelationship of Osteopontin, MMP-9 and ADAMTS4 in Patients With Osteoarthritis Undergoing Total Joint Arthroplasty.","date":"2020","source":"Clinical and applied thrombosis/hemostasis : official journal of the International Academy of Clinical and Applied Thrombosis/Hemostasis","url":"https://pubmed.ncbi.nlm.nih.gov/33350314","citation_count":24,"is_preprint":false},{"pmid":"26136925","id":"PMC_26136925","title":"Effect of osteopontin on the mRNA expression of ADAMTS4 and ADAMTS5 in chondrocytes from patients with knee osteoarthritis.","date":"2015","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/26136925","citation_count":24,"is_preprint":false},{"pmid":"11254106","id":"PMC_11254106","title":"Characterization of 5'-flanking region of human aggrecanase-1 (ADAMTS4) gene.","date":"2000","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/11254106","citation_count":22,"is_preprint":false},{"pmid":"22205175","id":"PMC_22205175","title":"Expression of ADAMTS-2, -3, -13, and -14 in culprit coronary lesions in patients with acute myocardial infarction or stable angina.","date":"2012","source":"Journal of thrombosis and thrombolysis","url":"https://pubmed.ncbi.nlm.nih.gov/22205175","citation_count":22,"is_preprint":false},{"pmid":"18671975","id":"PMC_18671975","title":"Calcium pentosan polysulfate directly inhibits enzymatic activity of ADAMTS4 (aggrecanase-1) in osteoarthritic chondrocytes.","date":"2008","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/18671975","citation_count":22,"is_preprint":false},{"pmid":"27082728","id":"PMC_27082728","title":"Cannabinoid WIN‑55,212‑2 mesylate inhibits ADAMTS‑4 activity in human osteoarthritic articular chondrocytes by inhibiting expression of syndecan‑1.","date":"2016","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/27082728","citation_count":21,"is_preprint":false},{"pmid":"29236314","id":"PMC_29236314","title":"IL-1β-induced miR-34a up-regulation inhibits Cyr61 to modulate osteoarthritis chondrocyte proliferation through ADAMTS-4.","date":"2018","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29236314","citation_count":21,"is_preprint":false},{"pmid":"16427204","id":"PMC_16427204","title":"Mammalian expression of full-length bovine aggrecan and link protein: formation of recombinant proteoglycan aggregates and analysis of proteolytic cleavage by ADAMTS-4 and MMP-13.","date":"2005","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/16427204","citation_count":21,"is_preprint":false},{"pmid":"22735305","id":"PMC_22735305","title":"Expression of ADAMTS-1, ADAMTS-4, ADAMTS-5 and TIMP3 by hepatocellular carcinoma cell lines.","date":"2012","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/22735305","citation_count":20,"is_preprint":false},{"pmid":"27844209","id":"PMC_27844209","title":"The impact of parathyroidectomy on serum ADAMTS1, ADAMTS4 levels, insulin resistance, and subclinical cardiovascular disease in primary hyperparathyroidism.","date":"2016","source":"Endocrine","url":"https://pubmed.ncbi.nlm.nih.gov/27844209","citation_count":20,"is_preprint":false},{"pmid":"16945513","id":"PMC_16945513","title":"Effects of covalently attached chondroitin sulfate on aggrecan cleavage by ADAMTS-4 and MMP-13.","date":"2006","source":"Matrix biology : journal of the International Society for Matrix Biology","url":"https://pubmed.ncbi.nlm.nih.gov/16945513","citation_count":20,"is_preprint":false},{"pmid":"36403758","id":"PMC_36403758","title":"Glucose-stimulated PGC-1α couples with CBP and Runx2 to mediate intervertebral disc degeneration through transactivation of ADAMTS4/5 in diet-induced obesity mice.","date":"2022","source":"Bone","url":"https://pubmed.ncbi.nlm.nih.gov/36403758","citation_count":19,"is_preprint":false},{"pmid":"21584905","id":"PMC_21584905","title":"Partially redundant functions of Adamts1 and Adamts4 in the perinatal development of the renal medulla.","date":"2011","source":"Developmental dynamics : an official publication of the American Association of Anatomists","url":"https://pubmed.ncbi.nlm.nih.gov/21584905","citation_count":19,"is_preprint":false},{"pmid":"16723216","id":"PMC_16723216","title":"An alternative spliced transcript of ADAMTS4 is present in human synovium from OA patients.","date":"2006","source":"Matrix biology : journal of the International Society for Matrix Biology","url":"https://pubmed.ncbi.nlm.nih.gov/16723216","citation_count":19,"is_preprint":false},{"pmid":"19342688","id":"PMC_19342688","title":"Adaptor proteins and Ras synergistically regulate IL-1-induced ADAMTS-4 expression in human chondrocytes.","date":"2009","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/19342688","citation_count":18,"is_preprint":false},{"pmid":"34718343","id":"PMC_34718343","title":"Synergistic upregulation of ADAMTS4 (aggrecanase-1) by cytokines and its suppression in knee osteoarthritic synovial fibroblasts.","date":"2021","source":"Laboratory investigation; a journal of technical methods and pathology","url":"https://pubmed.ncbi.nlm.nih.gov/34718343","citation_count":17,"is_preprint":false},{"pmid":"33300067","id":"PMC_33300067","title":"Inflammasome components and ADAMTS4 in premature rupture of membranes.","date":"2020","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/33300067","citation_count":17,"is_preprint":false},{"pmid":"28207788","id":"PMC_28207788","title":"Self-complementary adeno-associated virus serotype 6 mediated knockdown of ADAMTS4 induces long-term and effective enhancement of aggrecan in degenerative human nucleus pulposus cells: A new therapeutic approach for intervertebral disc disorders.","date":"2017","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/28207788","citation_count":17,"is_preprint":false},{"pmid":"20625753","id":"PMC_20625753","title":"Elevated level of ADAMTS4 in plasma and peripheral monocytes from patients with acute coronary syndrome.","date":"2010","source":"Clinical research in cardiology : official journal of the German Cardiac Society","url":"https://pubmed.ncbi.nlm.nih.gov/20625753","citation_count":16,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":57060,"output_tokens":9898,"usd":0.159825,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":20569,"output_tokens":8752,"usd":0.160823,"stage2_stop_reason":"end_turn"},"total_usd":0.320648,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2000,\n      \"finding\": \"ADAMTS4 (aggrecanase-1) cleaves aggrecan at the Glu373-Ala374 bond in the interglobular domain and at four additional sites within the chondroitin sulfate-rich region (KEEE1667-1668GLGS, GELE1480-1481GRGT, TAQE1771-1772AGEG, VSQE1871-1872LGQR), with cleavage in the CS-rich region occurring more efficiently than at the IGD site.\",\n      \"method\": \"In vitro cleavage assay with recombinant human ADAMTS4 on native aggrecan, followed by N-terminal sequence analysis of fragments\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct in vitro enzymatic assay with sequence-confirmed cleavage sites, replicated across multiple studies\",\n      \"pmids\": [\"10751421\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"The thrombospondin type-1 (TSP-1) motif of ADAMTS4 binds to glycosaminoglycans of aggrecan and is required for aggrecan substrate recognition and cleavage; truncated ADAMTS4 lacking the TSP-1 motif failed to cleave aggrecan, TSP-1 peptides blocked cleavage by competing with enzyme binding, and GAG-free aggrecan was not cleaved.\",\n      \"method\": \"Domain truncation mutagenesis, peptide competition assays, aggrecan cleavage assays with GAG-free aggrecan\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — multiple orthogonal in vitro approaches including mutagenesis and competition assays demonstrating substrate binding mechanism\",\n      \"pmids\": [\"10827174\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"ADAMTS4 cleaves brevican at the Glu395-Ser396 bond within the central non-homologous domain, a different site from that cleaved by MMPs (Ala360-Phe361), demonstrating substrate specificity distinct from MMP family members.\",\n      \"method\": \"In vitro digestion of brevican with purified ADAMTS4 and MMPs, followed by N-terminal sequence analysis of cleavage fragments\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct in vitro biochemical assay with sequence-confirmed cleavage sites\",\n      \"pmids\": [\"10986281\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"ADAMTS4 cleaves versican V1 at the Glu441-Ala442 bond, generating the DPEAAE neoepitope, as demonstrated by recombinant enzyme cleavage of recombinant and native human versican substrates; mature ADAMTS4 protein was also detected in aortic intima extracts.\",\n      \"method\": \"In vitro cleavage assay with recombinant ADAMTS4 and recombinant/native versican V1 substrates; Western analysis with neoepitope antisera; Western analysis of aortic tissue\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct in vitro enzymatic assay with sequence-confirmed cleavage site, combined with tissue detection\",\n      \"pmids\": [\"11278559\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"TIMP-3 is a potent inhibitor of ADAMTS4 (aggrecanase-1) with Ki values in the subnanomolar range; the N-terminal inhibitory domain of TIMP-3 alone is sufficient for this inhibition, whereas TIMP-1, TIMP-2 and TIMP-4 do not effectively inhibit ADAMTS4.\",\n      \"method\": \"In vitro inhibition kinetics assay with N-terminal TIMP-3 domain expressed from bacterial inclusion bodies\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — quantitative in vitro inhibition assay with recombinant proteins, replicated across multiple studies\",\n      \"pmids\": [\"11278243\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"ADAMTS4 activation requires C-terminal truncation from the p75 (or p68) form to the p60/p53 forms; only the p60 and p53 forms exhibit aggrecanase and versicanase activity; this truncation is mediated by a MMP (GPI-anchored type) and blocked by TIMP-1, MMP inhibitors, furin inhibitors, and inhibitors of GPI synthesis, but not by serine or cysteine protease inhibitors.\",\n      \"method\": \"Stable transfection of human chondrosarcoma cells with full-length ADAMTS4; domain-specific antisera Western analysis; preparative SDS-PAGE isolation of individual forms with activity assays; pharmacological inhibition studies\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods including fractionation, activity assays, and pharmacological dissection; replicated by other studies\",\n      \"pmids\": [\"11796708\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Full-length ADAMTS4 undergoes autocatalytic C-terminal truncation to generate ~53 kDa and ~40 kDa isoforms with reduced affinity for sulfated GAGs; the cysteine-rich and spacer domains contain additional GAG-binding sites beyond the thrombospondin motif, as demonstrated by competition with synthetic peptides mimicking GAG-binding consensus sequences.\",\n      \"method\": \"C-terminal sequencing and mass analysis of autocatalytic fragments; GAG-binding competition assays with deglycosylated aggrecan and synthetic peptides\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct biochemical characterization with sequence analysis and binding competition experiments\",\n      \"pmids\": [\"12202483\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"ADAMTS4 also cleaves the Asn341-Phe342 MMP site in aggrecan interglobular domain (secondarily to the Glu373-Ala374 aggrecanase site); this secondary cleavage is inhibited by TIMP-3 but not TIMP-1 or TIMP-2, distinguishing it from MMP activity.\",\n      \"method\": \"In vitro digestion of native human aggrecan and recombinant G1-G2 constructs with purified ADAMTS4; inhibitor studies with TIMP-1, TIMP-2, TIMP-3\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro enzymatic assay with mutagenesis of cleavage sites and selective inhibitor validation\",\n      \"pmids\": [\"11854269\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"The non-catalytic C-terminal spacer domain of ADAMTS4 masks its general proteolytic activity; deletion of the spacer domain generates a more promiscuous enzyme with enhanced activity against the Glu373-Ala374 IGD bond, Cm-Tf, fibromodulin, and decorin, while removal of the cysteine-rich domain reduces aggrecanase activity by 80%; the full-length 70 kDa form is the most effective aggrecanase but shows little IGD activity and binds to the pericellular matrix, whereas spacer-deleted ADAMTS4 is released from the matrix.\",\n      \"method\": \"Expression of domain deletion mutants in mammalian cells; aggrecanase activity assays; general proteinase substrate (Cm-Tf) assays; Western analysis with domain-specific antisera\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — systematic domain deletion mutagenesis with multiple substrate activity assays and localization studies\",\n      \"pmids\": [\"14662755\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Proprotein convertase furin cleaves the prodomain of pro-ADAMTS4 in the trans-Golgi network at multiple sites (RPRR206-209, RAKR209-212, or KR211-212), and the pro-form (but not the mature form) physically co-precipitates with furin; processing is blocked by brefeldin A and furin-specific inhibitors; a furin-independent activation pathway also exists.\",\n      \"method\": \"Co-localization by confocal microscopy; co-immunoprecipitation; RNA interference of furin; furin-specific inhibitor treatment; brefeldin A treatment\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-precipitation, RNAi, pharmacological inhibition, and localization in a single study\",\n      \"pmids\": [\"14744861\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"C-terminal truncation of ADAMTS4 from p68 to p53 (required for activation of IGD cleavage) is mediated by GPI-anchored MT4-MMP (MMP-17) on the cell surface; co-transfection with active MT4-MMP enhanced truncation, while inactive MT4-MMP mutant was ineffective; activated p53 ADAMTS4 remains associated with cell surface syndecan-1 through both chondroitin sulfate and heparan sulfate chains.\",\n      \"method\": \"Co-transfection of ADAMTS4 with active/inactive MT4-MMP; phosphatidylinositol-specific phospholipase C treatment; glycosaminoglycan lyase digestions with FACE analysis; immunoprecipitation with anti-syndecan-1\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods including active vs. inactive mutant controls, enzymatic digestion, and co-precipitation; independently supported by other studies on MT4-MMP/ADAMTS4 axis\",\n      \"pmids\": [\"14701864\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"ADAMTS4 binds to the C-terminal domain of fibronectin through its spacer domain; this interaction inhibits aggrecanase activity of ADAMTS4 (IC50 ~110 nM for full-length fibronectin; ~170 nM for 40-kDa C-terminal fibronectin fragment); ADAMTS4 lacking the spacer domain is not inhibited by fibronectin and does not co-localize with fibronectin on cell surfaces.\",\n      \"method\": \"Yeast two-hybrid screening; chemical cross-linking; solid-phase binding assay; confocal microscopy co-localization; aggrecanase activity assay with inhibition by fibronectin\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — yeast two-hybrid, cross-linking, solid-phase binding, and functional inhibition assay in a single study with domain mapping\",\n      \"pmids\": [\"15161923\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"ADAMTS4 cleaves versican V2 at Glu405-Gln406 to generate the brain protein GHAP (glial hyaluronate binding protein), as demonstrated by immunological analysis of purified human GHAP with anti-neoepitope antiserum and in vitro digestion of human cerebellum proteoglycans with ADAMTS4.\",\n      \"method\": \"Anti-neoepitope antiserum immunological analysis; in vitro digestion of human cerebellum proteoglycans with ADAMTS4; Western analysis\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical identification with neoepitope antibody and in vitro digestion, single lab\",\n      \"pmids\": [\"14561220\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Pro-ADAMTS4 is not capable of autocatalytic prodomain removal under physiological conditions (mercuric compounds, temperatures 22-65°C, ionic strengths 0.1-1M, neutral/acidic pH do not trigger autocatalysis); proprotein convertases furin, PACE4, and PC5/6 efficiently remove the prodomain at Arg212/Phe213 to generate active enzyme; MMP-9 and trypsin can also remove the prodomain; autocatalysis occurs only at basic pH 8-10.\",\n      \"method\": \"In vitro incubation of pro-ADAMTS4 with diverse proteases and conditions; activity assays; N-terminal sequencing\",\n      \"journal\": \"Archives of biochemistry and biophysics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — comprehensive in vitro assay screening multiple conditions with activity readout, single lab but systematic approach\",\n      \"pmids\": [\"16289022\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"MT4-MMP plays a central role in IL-1-induced cartilage aggrecanolysis by activating ADAMTS4; IL-1 treatment causes loss of p68 ADAMTS4 from cartilage and appearance of p53 in medium; blocking MT4-MMP (via ManN or EST) nearly completely prevented p68 loss, p53 formation, and aggrecan degradation; increased ADAMTS4/5 mRNA by IL-1 was not accompanied by increased total enzyme protein, indicating that activation rather than new synthesis drives aggrecanolysis.\",\n      \"method\": \"Western analysis of aggrecan fragments and ADAMTS4 species in cartilage explants; real-time PCR; pharmacological inhibition with ManN (GPI synthesis inhibitor) and EST (MMP expression inhibitor)\",\n      \"journal\": \"Osteoarthritis and cartilage\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple Western analyses and pharmacological inhibition in explant system, single lab\",\n      \"pmids\": [\"15780640\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"The spacer domain of ADAMTS4 is critical for its localization in the extracellular matrix, whereas for ADAMTS5 the cysteine-rich domain is more influential; sequential inclusion of C-terminal ancillary domains of ADAMTS4 progressively enhances activity against aggrecan, Cm-Tf, fibromodulin, decorin, biglycan, and fibronectin; very little proteolytic activity is detected with the catalytic domain alone.\",\n      \"method\": \"Expression of domain deletion mutants; activity assays against multiple substrates; ECM localization assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — systematic domain deletion mutagenesis with multiple substrate assays, comparative study with ADAMTS5 providing internal controls\",\n      \"pmids\": [\"17430884\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"The NFATp and Runx2 transcription factors regulate ADAMTS4 promoter activity in chondrocytes; the NFI binding site (-441 to -429) acts as a negative regulator of the ADAMTS4 promoter specifically in chondrocytes; the region -383 to +10 is necessary for full basal promoter activity and contains Sp1 and AP2 sites.\",\n      \"method\": \"Reporter gene (promoter-luciferase) assays with deletion variants; mRNA analysis in porcine chondrocytes and NIH3T3 cells\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — promoter reporter assays with deletion constructs in relevant cell types, single lab\",\n      \"pmids\": [\"16677612\", \"11254106\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"TIMP-3 inhibition of ADAMTS4 is enhanced in the presence of aggrecan through binding of chondroitin 6-sulfate GAGs to the thrombospondin type 1 motif and spacer domains of ADAMTS4, forming a complex with improved TIMP-3 binding affinity over free ADAMTS4.\",\n      \"method\": \"FRET peptide assay; aggrecan-binding competition experiments; solid-phase binding assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — quantitative FRET assay and binding studies, single lab, mechanistic dissection of substrate-inhibitor interplay\",\n      \"pmids\": [\"17470431\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Crystal structures of ADAMTS4 reveal two distinct catalytic site configurations: a closed autoinhibited non-binding form and an open binding-competent form; mature aggrecanases exist as an ensemble of at least two isomers, only one of which is proteolytically active.\",\n      \"method\": \"X-ray crystallography of human ADAMTS4 in apo and inhibitor-bound forms\",\n      \"journal\": \"Protein science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure determination, single study but direct structural evidence\",\n      \"pmids\": [\"18042673\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Aggrecan cleavage by ADAMTS4 proceeds via an exosite mechanism: substrate initially binds at an exosite (reflected in the apparent Km), then the peptide sequence binds at the active site; an active-site inhibitor (SC81956, hydroxamic acid, Ki=23 nM) is non-competitive with aggrecan but competitive with low-molecular-weight peptide substrates, consistent with inability to compete with the final Michaelis complex.\",\n      \"method\": \"Enzyme kinetics with native aggrecan and fluorogenic peptide substrates; inhibition kinetics with varying substrate concentrations\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — rigorous kinetic analysis with two substrate types, single lab\",\n      \"pmids\": [\"17487981\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Calcium pentosan polysulfate (CaPPS) directly inhibits the aggrecanase activity of ADAMTS4 by interacting with its C-terminal ancillary domains (thrombospondin type 1 repeat, cysteine-rich, and spacer domains), without affecting mRNA expression of ADAMTS species.\",\n      \"method\": \"Aggrecanase activity assay in IL-1α-stimulated OA chondrocytes; synthetic peptide competition of CaPPS binding; mRNA expression analysis\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct activity assay and domain mapping by peptide competition, single lab\",\n      \"pmids\": [\"18671975\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"ADAMTS4 processes the matricellular protein hevin in mouse brain; in vitro digestion of hevin with ADAMTS4 produced fragments similar to those present in brain lysates; a SPARC-like fragment generated from hevin co-localizes with ADAMTS4 in vivo in mouse cerebellum, and this proteolysis contributes to normal cerebellar development.\",\n      \"method\": \"In vitro digestion of hevin with recombinant ADAMTS4; co-localization by confocal microscopy with monoclonal antibodies; comparison of brain lysate fragments\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro cleavage assay combined with in vivo co-localization, single lab\",\n      \"pmids\": [\"20018883\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"ADAMTS4 is endocytosed and degraded by chondrocytes via LRP1; the cysteine-rich and spacer domains of ADAMTS4 are responsible for binding to LRP1 clusters II and IV (KD,app ~98 nM and ~73 nM, respectively); the half-life of ADAMTS4 endocytosis is ~220 min, slower than ADAMTS5 (~100 min) due to 13-fold lower affinity for LRP1; ADAMTS5 competitively inhibits ADAMTS4 endocytosis but not vice versa.\",\n      \"method\": \"Domain deletion mutagenesis; soluble LRP1 cluster binding assays; endocytosis kinetics; competitive inhibition assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — domain mutagenesis, quantitative binding affinity measurements, kinetic endocytosis assays, with clear mechanistic dissection\",\n      \"pmids\": [\"24474687\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"ADAMTS4 degrades brevican, neurocan, and phosphacan core proteins and reverses their inhibition of neurite outgrowth; local administration of ADAMTS4 protein promoted motor function recovery and enhanced axonal regeneration/sprouting after spinal cord contusion injury in mice.\",\n      \"method\": \"In vitro CSPG degradation assay; neurite outgrowth assay; in vivo spinal cord contusion model with local ADAMTS4 administration and behavioral assessment\",\n      \"journal\": \"Journal of neuroinflammation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro substrate degradation combined with in vivo functional assay, single lab\",\n      \"pmids\": [\"22420304\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"ADAMTS4 and ADAMTS5 participate in regulated proteolytic processing of Reelin; both enzymes cleave Reelin in vitro, and TIMP-3, α-2-Macroglobulin, serpins, and MMP-9 modulate this cleavage; ADAMTS4 and Reelin expression levels largely overlap in murine hippocampus.\",\n      \"method\": \"In vitro cleavage assay with recombinant enzymes and Reelin; inhibitor studies with TIMPs and serpins; immunolocalization in murine hippocampus\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct in vitro cleavage assay with inhibitor dissection, combined with in vivo localization, single lab\",\n      \"pmids\": [\"23082219\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"TNF-α and IL-1β regulate ADAMTS4 expression in nucleus pulposus cells through MAPK (ERK1, p38α, p38β2, p38γ) and NF-κB signaling; p65 induces ADAMTS4 promoter activity while p50 blocks it; silencing of NF-κB components (p65, p52, IKK-α, IKK-β) decreased ADAMTS-4 and -5 levels and aggrecan degradation; both ADAMTS-4 and ADAMTS-5 contribute non-redundantly to aggrecan degradation in human NP cells.\",\n      \"method\": \"Transient transfection promoter assays; MAPK/NF-κB inhibitors; gain/loss-of-function studies; lentiviral shRNA silencing; Western blotting; aggrecan degradation assays\",\n      \"journal\": \"The American journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple pathway inhibitors and genetic knockdown with functional readout, single lab\",\n      \"pmids\": [\"23602832\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"IL-1β-induced upregulation of ADAMTS4 in human OA chondrocytes is suppressed by miR-125b overexpression; luciferase reporter assay with mutated miR-125b binding site in the ADAMTS4 3'UTR confirmed direct interaction, establishing miR-125b as a direct post-transcriptional regulator of ADAMTS4.\",\n      \"method\": \"miR-125b overexpression; luciferase reporter assay with wild-type and mutant ADAMTS4 3'UTR; quantitative RT-PCR; Western blotting\",\n      \"journal\": \"Arthritis research & therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — luciferase assay with mutation validation plus functional overexpression experiment, single lab\",\n      \"pmids\": [\"23406982\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CCN1 (Cyr61) binds specifically to the cysteine-rich domain of ADAMTS4 and inhibits its aggrecanase activity; CCN1 co-purifies with ADAMTS4-transfected chondrocytic cells (identified by LC-MS/MS), and the interaction was confirmed by immunoprecipitation and solid-phase binding assay; in TGFβ-treated chondrocytes, CCN1 knockdown reveals latent ADAMTS4 aggrecanase activity.\",\n      \"method\": \"LC-MS/MS identification of co-purified proteins; immunoprecipitation; solid-phase binding assay; aggrecan digestion assay; siRNA knockdown; immunohistochemistry\",\n      \"journal\": \"Arthritis & rheumatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple binding assays combined with functional inhibition and genetic knockdown, single lab\",\n      \"pmids\": [\"25709087\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"ADAMTS4 translocates to the nucleus in smooth muscle cells (SMCs) under stress and directly cleaves and degrades poly ADP ribose polymerase-1 (PARP-1), leading to SMC apoptosis; ADAMTS4 deficiency in mice significantly reduced angiotensin II/high-fat diet-induced aortic aneurysm formation, elastic fiber destruction, versican degradation, macrophage infiltration, and apoptosis.\",\n      \"method\": \"Adamts4-/- mouse model of sporadic AAD (angiotensin II + high fat diet); nuclear translocation demonstrated by imaging of apoptotic SMCs; direct PARP-1 cleavage assay; aortic phenotyping (diameter, histology)\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — knockout mouse model combined with direct cleavage assay, single lab; nuclear localization is notable but mechanistic evidence for PARP-1 cleavage relies on single study\",\n      \"pmids\": [\"28955046\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"ADAMTS4 cleaves APP/Aβ to generate N-truncated Aβ4-x peptides; a recognition site for ADAMTS4 was identified in the Aβ sequence; inducible overexpression of ADAMTS4 in HEK293 cells increased secretion of Aβ4-40; in ADAMTS4-/- mice (5xFAD background) Aβ4-40 levels were reduced; ADAMTS4 is exclusively expressed in oligodendrocytes in adult mouse brain, and ADAMTS4-/- oligodendrocyte cultures do not produce Aβ4-40.\",\n      \"method\": \"ADAMTS4 recognition site identification; inducible overexpression in HEK293 cells with Aβ ELISA; ADAMTS4-/- mouse model; primary oligodendrocyte cultures from ADAMTS4-/- mice; immunofluorescence co-localization\",\n      \"journal\": \"Acta neuropathologica\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — overexpression gain-of-function, knockout loss-of-function, and cell-specific culture experiments with consistent results, single lab but multiple orthogonal approaches\",\n      \"pmids\": [\"30426203\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Sox4 (and Sox11) directly bind to the ADAMTS4 promoter and upregulate ADAMTS4 and ADAMTS5 gene expression; demonstrated by luciferase reporter assay and chromatin immunoprecipitation; Sox4 overexpression in mouse femoral head cartilage organ cultures caused cartilage destruction associated with increased aggrecanase expression.\",\n      \"method\": \"Luciferase reporter assay; chromatin immunoprecipitation (ChIP); adenoviral overexpression in mouse femoral head organ cultures; quantitative RT-PCR\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and reporter assay confirm direct promoter binding with functional validation in organ culture, single lab\",\n      \"pmids\": [\"30016600\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Damage-responsive lung fibroblasts produce ADAMTS4 during severe respiratory viral infection; ADAMTS4 ECM protease activity modifies the lung microenvironment to promote immune cell infiltration; ADAMTS4 levels in lower respiratory tract samples correlated with severity of influenza infection in three human cohorts.\",\n      \"method\": \"Single-cell transcriptomics identifying fibroblast activation states; mouse influenza model with fibroblast characterization; measurement of ADAMTS4 in human BALF from influenza patients\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mechanistic identification of ADAMTS4-producing fibroblast subpopulation with human cohort validation, but specific molecular mechanism of immune cell infiltration promotion is correlative\",\n      \"pmids\": [\"33116313\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ADAMTS4 (along with ADAMTS1 and ADAMTS5) cleaves versican V1 at multiple sites beyond the known Glu441-Ala442 bond; 21 novel cleavage sites were identified using LC-MS/MS label-free proteomics with z-score ranking; ADAMTS4 shows a substrate site preference for P1-Glu residue.\",\n      \"method\": \"In vitro digestion of recombinant versican V1 with recombinant full-length ADAMTS4 vs. catalytically inactive mutant; LC-MS/MS label-free quantitative proteomics with z-score analysis\",\n      \"journal\": \"Journal of proteomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — systematic in vitro enzymatic assay with inactive mutant controls and quantitative proteomics, single lab\",\n      \"pmids\": [\"34450332\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Adamts4 and Adamts1 play redundant roles in perinatal kidney development; Adamts4 single-knockout mice are phenotypically normal, but >95% of Adamts1-/-;Adamts4-/- double-knockout mice die within 72 hours of birth with marked thinning of the renal medulla, a defect not observed in embryos.\",\n      \"method\": \"Gene targeting to generate Adamts4-/- mice; genetic cross with Adamts1-/- mice; histological analysis of renal medulla\",\n      \"journal\": \"Developmental dynamics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis using double-knockout mouse model with clear anatomical phenotype, single lab\",\n      \"pmids\": [\"21584905\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"IL-1β induction of ADAMTS4 expression in human OA chondrocytes requires PKCζ (an atypical PKC); PKCζ mediates IL-1β-induced NF-κB activation (phosphorylation of IKKαβ and IκBα); pharmacological inhibition or siRNA/shRNA knockdown of PKCζ suppressed IL-1β-induced ADAMTS4 mRNA upregulation and aggrecanase activity.\",\n      \"method\": \"Pharmacological inhibitors of atypical PKCs; siRNA and shRNA knockdown of PKCζ; Western blotting for NF-κB pathway phosphorylation; quantitative PCR for ADAMTS4 mRNA; aggrecanase activity assay\",\n      \"journal\": \"Arthritis and rheumatism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knockdown plus pharmacological inhibition with pathway readout and functional assay, single lab\",\n      \"pmids\": [\"18050214\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"IL-1β induction of ADAMTS4 in chondrocytes requires MyD88, IRAK1, and TRAF6 adaptor proteins (each individually required but partial); Ras-mediated ROS production synergizes with these adaptors; combined knockdown of Ras and individual adaptors strongly blocked NF-κB activation (IKKαβ, IκBα phosphorylation) and ADAMTS4 induction.\",\n      \"method\": \"siRNA-mediated knockdown of MyD88, IRAK1, TRAF6, and Ras; antioxidant treatment; Western blotting for NF-κB pathway; ADAMTS4 mRNA quantitation\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic siRNA knockdown with mechanistic pathway readout, single lab\",\n      \"pmids\": [\"19342688\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"ADAMTS4 exerts anti-inflammatory effects in microglia and astrocytes; recombinant ADAMTS4 pretreatment of primary microglia and astrocytes decreased LPS-induced NO production and pro-inflammatory cytokine synthesis/release (NOS2, CCL2, TNF-α, IL-1β, MMP-9); siRNA silencing of ADAMTS4 increased these pro-inflammatory markers; in vivo, ADAMTS4 treatment decreased astrogliosis, macrophage infiltration, and increased M2 microglia after middle cerebral artery occlusion.\",\n      \"method\": \"Recombinant ADAMTS4 treatment of primary microglia/astrocyte cultures; siRNA silencing of ADAMTS4; NO and cytokine measurement; mouse middle cerebral artery occlusion model with ADAMTS4 treatment; immunohistochemistry\",\n      \"journal\": \"Glia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain- and loss-of-function in vitro with in vivo validation, single lab\",\n      \"pmids\": [\"27301579\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"ADAMTS4 contributes to aggrecan degradation in human osteoarthritic nucleus pulposus cells non-redundantly with ADAMTS5; silencing either enzyme individually reduced TNF-α-dependent aggrecan degradation.\",\n      \"method\": \"Lentiviral shRNA silencing of ADAMTS-4 or ADAMTS-5 in human NP cells; Western blotting; aggrecan degradation assay\",\n      \"journal\": \"The American journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — lentiviral knockdown of individual genes with functional aggrecan degradation readout, single lab\",\n      \"pmids\": [\"23602832\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ADAMTS4 is a secreted, multi-domain metalloprotease (containing catalytic, disintegrin, thrombospondin, cysteine-rich, and spacer domains) that is synthesized as a latent zymogen, activated by furin/proprotein convertases in the trans-Golgi network and by GPI-anchored MT4-MMP-mediated C-terminal truncation on the cell surface; the activated enzyme cleaves aggrecan at multiple sites (predominantly in the chondroitin sulfate-rich region via exosite-mediated GAG binding through its TSP-1 motif and C-terminal domains), as well as versican, brevican, neurocan, hevin, Reelin, fibulin-2, and APP/Aβ; its activity is regulated by TIMP-3 (potent inhibitor), fibronectin (C-terminal domain inhibitor), CCN1 (cysteine-rich domain inhibitor), LRP1-mediated endocytosis, and transcriptionally by NF-κB, MAPK pathways, furin convertases, Runx2, NFATp, Sox4, and miR-125b; C-terminal spacer domain deletion releases the enzyme from the pericellular matrix and broadens its substrate specificity, while syndecan-1 anchors the activated p53 form to the cell surface.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ADAMTS4 (aggrecanase-1) is a secreted, multidomain metalloprotease that remodels the extracellular and pericellular matrix by cleaving large chondroitin sulfate proteoglycans, most prominently aggrecan, which it cuts at the Glu373-Ala374 interglobular-domain bond and at multiple more efficiently processed sites in the chondroitin sulfate-rich region [#0, #7]. It also cleaves versican (V1 at Glu441-Ala442 and many additional sites, V2 to generate brain GHAP), brevican, and other CNS proteoglycans, establishing distinct roles in connective tissue and neural matrix turnover [#2, #3, #12, #23, #32]. Substrate recognition and activity depend critically on its C-terminal ancillary domains: the thrombospondin type-1 motif and cysteine-rich/spacer domains bind glycosaminoglycans to position substrate, sequential inclusion of these domains progressively enhances proteolysis, and aggrecan cleavage proceeds through an exosite mechanism in which substrate first docks at an exosite before the scissile peptide enters the active site [#1, #6, #15, #19]. The enzyme is produced as a latent zymogen activated by furin/proprotein convertase removal of the prodomain in the trans-Golgi and by GPI-anchored MT4-MMP (MMP-17)-mediated C-terminal truncation at the cell surface, where the active form is retained via syndecan-1; this activation step, rather than new synthesis, drives IL-1-induced cartilage aggrecanolysis [#5, #9, #10, #14]. The spacer domain confines full-length enzyme to the pericellular matrix and masks broad proteolytic activity, so its removal releases the enzyme and broadens substrate specificity [#8, #11]. ADAMTS4 activity is restrained by TIMP-3 (a potent inhibitor whose efficacy is enhanced by aggrecan GAGs), fibronectin (via the spacer domain), and CCN1 (via the cysteine-rich domain), and the enzyme is cleared by LRP1-mediated endocytosis [#4, #11, #17, #22, #27]. Transcriptionally it is driven by inflammatory cytokine signaling through PKCζ, MyD88/IRAK1/TRAF6, MAPK and NF-κB pathways and by Runx2, NFATp, and Sox4, and is repressed post-transcriptionally by miR-125b [#25, #26, #30, #34, #35]. Beyond matrix remodeling, ADAMTS4 cleaves APP/Aβ to generate N-truncated Aβ4-x peptides in oligodendrocytes, translocates to the nucleus of stressed smooth muscle cells to degrade PARP-1 and promote apoptosis in aortic aneurysm, and modulates neuroinflammation and CNS injury responses [#28, #29, #36].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Established the defining enzymatic identity of ADAMTS4 as aggrecanase-1 by mapping the precise aggrecan bonds it cleaves, distinguishing aggrecanase from MMP-type cleavage.\",\n      \"evidence\": \"In vitro cleavage of native aggrecan with recombinant ADAMTS4 and N-terminal sequencing of fragments\",\n      \"pmids\": [\"10751421\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not address how non-catalytic domains contribute to site selection\", \"Cleavage efficiency in cell/tissue context not yet defined\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Showed that substrate recognition is not purely active-site-driven but requires GAG-mediated binding through the thrombospondin type-1 motif, defining an ancillary-domain recognition mechanism.\",\n      \"evidence\": \"Domain truncation mutagenesis, TSP-1 peptide competition, and assays with GAG-free aggrecan\",\n      \"pmids\": [\"10827174\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Contributions of cysteine-rich and spacer domains to GAG binding not yet resolved\", \"Structural basis of exosite binding not defined\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Extended the substrate range beyond aggrecan to brevican, showing ADAMTS4 cleaves CNS proteoglycans at sites distinct from MMPs.\",\n      \"evidence\": \"In vitro digestion of brevican with purified ADAMTS4 and MMPs with N-terminal sequencing\",\n      \"pmids\": [\"10986281\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance of brevican cleavage not established\", \"Cell type producing the enzyme in brain not identified\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Identified TIMP-3 as the selective endogenous inhibitor of ADAMTS4, localizing the inhibitory activity to the TIMP-3 N-terminal domain.\",\n      \"evidence\": \"In vitro inhibition kinetics with recombinant TIMP domains\",\n      \"pmids\": [\"11278243\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How TIMP-3 engages the catalytic site structurally not resolved\", \"Physiological regulation of TIMP-3/ADAMTS4 balance in tissue untested\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Demonstrated versican as a physiological substrate and detected mature ADAMTS4 in vascular tissue, broadening its role to connective tissue beyond cartilage.\",\n      \"evidence\": \"In vitro versican cleavage with neoepitope antisera plus Western analysis of aortic intima\",\n      \"pmids\": [\"11278559\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of vascular versican cleavage not tested at this stage\", \"Activation state of the detected tissue enzyme unknown\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Defined the activation requirement for ADAMTS4: only C-terminally truncated p60/p53 forms are proteolytically active, and truncation is mediated by a GPI-anchored MMP, separating zymogen maturation from full enzymatic competence.\",\n      \"evidence\": \"Stable expression in chondrosarcoma cells, form fractionation with activity assays, and pharmacological inhibition\",\n      \"pmids\": [\"11796708\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the GPI-anchored MMP not yet established\", \"Relationship of truncation to substrate specificity not defined\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Established that C-terminal ancillary domains carry GAG-binding sites and that autocatalytic truncation reduces GAG affinity, linking domain composition to matrix anchoring.\",\n      \"evidence\": \"C-terminal sequencing of autocatalytic fragments and GAG-binding competition with synthetic peptides\",\n      \"pmids\": [\"12202483\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological trigger for autocatalysis not defined\", \"Quantitative impact on substrate cleavage in tissue not measured\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Showed ADAMTS4 also cleaves the aggrecan MMP-site secondarily, and that this is TIMP-3-sensitive, reinforcing its mechanistic distinction from MMPs.\",\n      \"evidence\": \"In vitro digestion of aggrecan and G1-G2 constructs with selective TIMP inhibitors\",\n      \"pmids\": [\"11854269\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Biological significance of the secondary MMP-site cleavage unclear\", \"Order/kinetics relative to primary cleavage not resolved in vivo\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Revealed that the spacer domain masks general proteolytic activity and confines the enzyme to the pericellular matrix, so domain removal expands substrate range and releases the enzyme.\",\n      \"evidence\": \"Domain deletion mutants assayed against multiple substrates with localization analysis\",\n      \"pmids\": [\"14662755\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo prevalence of spacer-deleted forms unknown\", \"Mechanism of substrate masking at structural level not resolved here\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Identified furin as the trans-Golgi prodomain-processing protease for pro-ADAMTS4 while noting a parallel furin-independent route.\",\n      \"evidence\": \"Co-localization, reciprocal co-IP, furin RNAi, and inhibitor/brefeldin A treatment\",\n      \"pmids\": [\"14744861\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the furin-independent pathway not defined\", \"Whether prodomain removal alone confers activity not fully separated from C-terminal events\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Identified MT4-MMP (MMP-17) as the GPI-anchored protease that truncates ADAMTS4 to the active p53 form at the cell surface, where syndecan-1 retains it.\",\n      \"evidence\": \"Co-transfection with active/inactive MT4-MMP, PI-PLC and GAG lyase treatments, and anti-syndecan-1 co-IP\",\n      \"pmids\": [\"14701864\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Regulation of MT4-MMP availability in tissue not addressed\", \"Whether other surface proteases substitute in vivo unknown\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Defined fibronectin as a spacer-domain-dependent inhibitor and co-localization partner, adding a matrix-based brake on aggrecanase activity.\",\n      \"evidence\": \"Yeast two-hybrid, cross-linking, solid-phase binding, and inhibition assays with domain mapping\",\n      \"pmids\": [\"15161923\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance of fibronectin inhibition not tested\", \"Competition with substrate GAG binding not quantified\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Linked ADAMTS4 to generation of the brain proteoglycan fragment GHAP via versican V2 cleavage, reinforcing a CNS matrix role.\",\n      \"evidence\": \"Anti-neoepitope analysis of purified GHAP and in vitro digestion of cerebellar proteoglycans\",\n      \"pmids\": [\"14561220\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo enzyme responsible not genetically confirmed\", \"Functional consequence of GHAP generation untested\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Clarified the biochemical constraints on activation, showing prodomain removal under physiological conditions requires proprotein convertases (furin, PACE4, PC5/6) rather than autocatalysis.\",\n      \"evidence\": \"In vitro incubation of pro-ADAMTS4 with multiple proteases across conditions with activity assays\",\n      \"pmids\": [\"16289022\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative in vivo contribution of each convertase not defined\", \"Integration with C-terminal MT4-MMP step not resolved\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Demonstrated in cartilage explants that IL-1-driven aggrecanolysis depends on MT4-MMP-mediated ADAMTS4 activation rather than increased enzyme synthesis.\",\n      \"evidence\": \"Western analysis of ADAMTS4 species and aggrecan fragments with GPI/MMP pharmacological inhibition in explants\",\n      \"pmids\": [\"15780640\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab explant system; genetic confirmation absent\", \"Distinction from ADAMTS5 contribution incomplete\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Mapped the transcriptional control of the ADAMTS4 promoter in chondrocytes, identifying NFATp, Runx2 and a chondrocyte-specific NFI repressor element.\",\n      \"evidence\": \"Promoter-luciferase deletion reporter assays and mRNA analysis in chondrocytes\",\n      \"pmids\": [\"16677612\", \"11254106\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct factor binding (e.g., ChIP) not shown for all sites\", \"Cytokine-responsive elements not yet integrated\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Systematized the domain hierarchy controlling activity and localization, showing sequential ancillary domains progressively enhance proteolysis and that the spacer domain governs ECM localization (versus cysteine-rich for ADAMTS5).\",\n      \"evidence\": \"Comparative domain-deletion mutagenesis with multi-substrate assays against ADAMTS5\",\n      \"pmids\": [\"17430884\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural mechanism of domain cooperation not resolved here\", \"In vivo consequences of domain configuration untested\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Provided structural and kinetic mechanisms: crystal structures revealed open/closed catalytic configurations, and kinetics defined an exosite-mediated aggrecan cleavage with implications for inhibitor design.\",\n      \"evidence\": \"X-ray crystallography of apo/inhibitor-bound enzyme; enzyme kinetics with aggrecan and peptide substrates plus active-site inhibitor\",\n      \"pmids\": [\"18042673\", \"17487981\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Substrate-bound structure not solved\", \"Physiological trigger switching open/closed states unknown\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Showed aggrecan GAG binding enhances TIMP-3 inhibition, revealing substrate-inhibitor interplay at the ancillary domains.\",\n      \"evidence\": \"FRET peptide assay with aggrecan-binding competition and solid-phase binding\",\n      \"pmids\": [\"17470431\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab mechanistic study; in vivo relevance untested\", \"Structural basis of the ternary complex unresolved\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Identified upstream cytokine signaling requirements (PKCζ, MyD88/IRAK1/TRAF6, Ras/ROS) for IL-1β-driven ADAMTS4 induction via NF-κB.\",\n      \"evidence\": \"siRNA/shRNA knockdown and pharmacological inhibition with NF-κB pathway and ADAMTS4 mRNA readouts in chondrocytes\",\n      \"pmids\": [\"18050214\", \"19342688\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Promoter elements engaged by these pathways not fully mapped\", \"Single-lab studies; in vivo validation limited\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Extended substrate range to the matricellular protein hevin and linked ADAMTS4 proteolysis to cerebellar development in vivo.\",\n      \"evidence\": \"In vitro hevin digestion and in vivo co-localization in mouse cerebellum\",\n      \"pmids\": [\"20018883\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Genetic loss-of-function effect on hevin in vivo not shown\", \"Functional outcome of the SPARC-like fragment unresolved\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Revealed a developmental role through genetic redundancy: Adamts4 alone is dispensable, but Adamts1/Adamts4 double knockouts die perinatally with renal medulla defects.\",\n      \"evidence\": \"Gene targeting and genetic cross with histological renal analysis\",\n      \"pmids\": [\"21584905\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular substrate underlying the renal phenotype unidentified\", \"Extent of redundancy with other ADAMTS members untested\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Established LRP1-mediated endocytosis as the clearance route for ADAMTS4 via cysteine-rich/spacer binding, with slower turnover than ADAMTS5.\",\n      \"evidence\": \"Domain mutagenesis, soluble LRP1 cluster binding affinity, and endocytosis kinetics with competition assays\",\n      \"pmids\": [\"24474687\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo contribution of LRP1 clearance to matrix turnover not quantified\", \"Regulation of LRP1 availability not addressed\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Demonstrated CNS therapeutic relevance: ADAMTS4 degrades inhibitory CSPGs (brevican, neurocan, phosphacan) and promotes axonal regeneration and motor recovery after spinal cord injury.\",\n      \"evidence\": \"In vitro CSPG degradation and neurite outgrowth assays plus in vivo spinal cord contusion model\",\n      \"pmids\": [\"22420304\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Endogenous versus exogenous enzyme contribution not separated\", \"Long-term and mechanistic basis of recovery not fully defined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Added Reelin to the substrate repertoire under multi-inhibitor control, connecting ADAMTS4 to hippocampal Reelin processing.\",\n      \"evidence\": \"In vitro cleavage with inhibitor dissection and hippocampal immunolocalization\",\n      \"pmids\": [\"23082219\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo Reelin processing by ADAMTS4 genetically unconfirmed\", \"Functional consequence for Reelin signaling untested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defined cytokine-driven transcriptional control in nucleus pulposus cells through MAPK/NF-κB and showed ADAMTS4 contributes non-redundantly to aggrecan degradation alongside ADAMTS5.\",\n      \"evidence\": \"Promoter assays, pathway inhibitors, lentiviral shRNA silencing, and aggrecan degradation assays\",\n      \"pmids\": [\"23602832\", \"23406982\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"miR-125b regulation shown by reporter, not in vivo\", \"Relative ADAMTS4/ADAMTS5 contribution context-dependent\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified CCN1 as a cysteine-rich-domain-binding inhibitor that keeps ADAMTS4 latent, revealing an additional endogenous brake unmasked by CCN1 knockdown.\",\n      \"evidence\": \"LC-MS/MS co-purification, immunoprecipitation, solid-phase binding, aggrecan digestion, and siRNA knockdown\",\n      \"pmids\": [\"25709087\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of CCN1 inhibition unresolved\", \"In vivo relevance to OA progression untested\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Uncovered an anti-inflammatory role in CNS glia, where ADAMTS4 suppresses microglial/astrocyte pro-inflammatory responses and improves outcome after cerebral ischemia.\",\n      \"evidence\": \"Recombinant treatment and siRNA silencing in glia with cytokine measurement and a middle cerebral artery occlusion model\",\n      \"pmids\": [\"27301579\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism of anti-inflammatory action undefined\", \"Substrate mediating glial effects unidentified\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Revealed a non-canonical intracellular function: stress-induced nuclear translocation and PARP-1 cleavage driving smooth muscle apoptosis in aortic aneurysm/dissection.\",\n      \"evidence\": \"Adamts4-/- mouse aneurysm model, nuclear translocation imaging, and direct PARP-1 cleavage assay\",\n      \"pmids\": [\"28955046\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism of nuclear import for a secreted protease unexplained\", \"PARP-1 cleavage relies on a single study\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established ADAMTS4 as an oligodendrocyte-specific generator of N-truncated Aβ4-x peptides, implicating it in amyloid processing.\",\n      \"evidence\": \"Recognition-site mapping, inducible overexpression with Aβ ELISA, ADAMTS4-/- mice, and ADAMTS4-/- oligodendrocyte cultures\",\n      \"pmids\": [\"30426203\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Contribution to amyloid pathology in human disease not established\", \"Regulation of oligodendrocyte ADAMTS4 expression unclear\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Identified Sox4 (and Sox11) as direct transcriptional activators of ADAMTS4 promoting cartilage destruction.\",\n      \"evidence\": \"Luciferase reporter, ChIP, and adenoviral overexpression in femoral head organ cultures\",\n      \"pmids\": [\"30016600\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Integration with cytokine-driven regulation not mapped\", \"In vivo Sox4-ADAMTS4 axis in OA untested\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Linked ADAMTS4 to acute infection biology, showing damage-responsive lung fibroblasts produce it to remodel ECM and promote immune infiltration, with levels tracking influenza severity.\",\n      \"evidence\": \"Single-cell transcriptomics, mouse influenza model, and human BALF measurements across cohorts\",\n      \"pmids\": [\"33116313\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Specific ECM substrate driving immune infiltration not defined\", \"Causal mechanism in humans remains correlative\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Refined substrate-site preference by mapping 21 novel versican V1 cleavage sites and a P1-Glu preference, expanding the proteolytic landscape.\",\n      \"evidence\": \"In vitro versican digestion with active versus inactive enzyme and label-free LC-MS/MS proteomics\",\n      \"pmids\": [\"34450332\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Biological significance of individual novel sites untested\", \"In vivo occurrence of these cleavages not confirmed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How a secreted matrix protease achieves nuclear localization and intracellular PARP-1 cleavage, and how its diverse extracellular and intracellular activities are coordinated across tissues, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No mechanism for nuclear import of ADAMTS4 established\", \"Integration of matrix-remodeling, anti-inflammatory, and apoptotic functions not unified\", \"Substrate driving most in vivo phenotypes often unidentified\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 2, 3, 21, 28, 29, 32]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0, 5, 13]},\n      {\"term_id\": \"GO:0140098\", \"supporting_discovery_ids\": [1, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005576\", \"supporting_discovery_ids\": [3, 5, 31]},\n      {\"term_id\": \"GO:0031012\", \"supporting_discovery_ids\": [8, 11, 15]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [10, 5]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [9]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [28]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1474244\", \"supporting_discovery_ids\": [0, 3, 8]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [5, 9, 13]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [28, 29, 31]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"TIMP3\", \"MMP17\", \"SDC1\", \"FN1\", \"CCN1\", \"LRP1\", \"FURIN\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}