{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,5]],"date-time":"2026-11-05T04:31:28Z","timestamp":1793853088226,"version":"4.6.1"},"reference-count":46,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,6,14]],"date-time":"2023-06-14T00:00:00Z","timestamp":1686700800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007074","name":"Fondazione CON IL SUD","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100007074","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Mol. Biosci."],"abstract":"<jats:p>ADAM15 is a member of the disintegrin-metalloproteinase family of sheddases, which plays a role in several biological processes including cartilage homeostasis. In contrast with well-characterized ADAMs, such as the canonical sheddases ADAM17 and ADAM10, little is known about substrates of ADAM15 or how the enzyme exerts its biological functions. Herein, we used \u201csurface-spanning enrichment with click-sugars (SUSPECS)\u201d proteomics to identify ADAM15 substrates and\/or proteins regulated by the proteinase at the cell surface of chondrocyte-like cells. Silencing of ADAM15 by siRNAs significantly altered membrane levels of 13 proteins, all previously not known to be regulated by ADAM15. We used orthogonal techniques to validate ADAM15 effects on 3 of these proteins which have known roles in cartilage homeostasis. This confirmed that ADAM15-silencing increased cell surface levels of the programmed cell death 1 ligand 2 (PDCD1LG2) and reduced cell surface levels of vasorin and the sulfate transporter SLC26A2 through an unknown post-translational mechanism. The increase of PDCD1LG2 by ADAM15 knockdown, a single-pass type I transmembrane protein, suggested it could be a proteinase substrate. However, shed PDCD1LG2 could not be detected even by a data-independent acquisition mass spectrometry, a highly sensitive method for identification and quantification of proteins in complex protein samples, suggesting that ADAM15 regulates PDCD1LG2 membrane levels by a mechanism different from ectodomain shedding.<\/jats:p>","DOI":"10.3389\/fmolb.2023.1162504","type":"journal-article","created":{"date-parts":[[2023,6,14]],"date-time":"2023-06-14T09:23:09Z","timestamp":1686734589000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Identification of membrane proteins regulated by ADAM15 by SUSPECS proteomics"],"prefix":"10.3389","volume":"10","author":[{"given":"Matteo","family":"Calligaris","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chun Y.","family":"Yang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simone","family":"Bonelli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donatella Pia","family":"Span\u00f2","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephan A.","family":"M\u00fcller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan F.","family":"Lichtenthaler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linda","family":"Troeberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simone D.","family":"Scilabra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,6,14]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1946","DOI":"10.1002\/1529-0131(199909)42:9<1946::AID-ANR21>3.0.CO;2-E","article-title":"Up-regulation of MDC15 (metargidin) messenger RNA in human osteoarthritic cartilage","volume":"42","author":"Bohm","year":"1999","journal-title":"Arthritis Rheum."},{"key":"B2","doi-asserted-by":"publisher","first-page":"1100","DOI":"10.1002\/art.20974","article-title":"Homeostatic effects of the metalloproteinase disintegrin ADAM15 in degenerative cartilage remodeling","volume":"52","author":"Bohm","year":"2005","journal-title":"Arthritis Rheum."},{"key":"B3","doi-asserted-by":"publisher","first-page":"1372","DOI":"10.1002\/art.27387","article-title":"ADAM15 exerts an antiapoptotic effect on osteoarthritic chondrocytes via up-regulation of the X-linked inhibitor of apoptosis","volume":"62","author":"Bohm","year":"2010","journal-title":"Arthritis Rheum."},{"key":"B4","doi-asserted-by":"publisher","first-page":"641","DOI":"10.1096\/fj.07-8876rev","article-title":"ADAM-15: A metalloprotease that mediates inflammation","volume":"22","author":"Charrier-Hisamuddin","year":"2008","journal-title":"FASEB J."},{"key":"B5","doi-asserted-by":"publisher","first-page":"3278","DOI":"10.1002\/art.34566","article-title":"Chondrocyte-intrinsic Smad3 represses Runx2-inducible matrix metalloproteinase 13 expression to maintain articular cartilage and prevent osteoarthritis","volume":"64","author":"Chen","year":"2012","journal-title":"Arthritis Rheum."},{"key":"B6","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1016\/j.matbio.2021.12.002","article-title":"ADAM15 is required for optimal collagen cross-linking and scar formation following myocardial infarction","volume":"105","author":"Chute","year":"2022","journal-title":"Matrix Biol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"1697","DOI":"10.1242\/dev.030742","article-title":"Sulfation of chondroitin sulfate proteoglycans is necessary for proper Indian hedgehog signaling in the developing growth plate","volume":"136","author":"Cortes","year":"2009","journal-title":"Development"},{"key":"B8","doi-asserted-by":"publisher","first-page":"L325","DOI":"10.1152\/ajplung.00294.2014","article-title":"ADAM-Family metalloproteinases in lung inflammation: Potential therapeutic targets","volume":"308","author":"Dreymueller","year":"2015","journal-title":"Am. J. Physiol. Lung Cell Mol. Physiol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"991","DOI":"10.3892\/mmr.2013.1272","article-title":"ADAM15 is involved in MICB shedding and mediates the effects of gemcitabine on MICB shedding in PANC-1 pancreatic cancer cells","volume":"7","author":"Duan","year":"2013","journal-title":"Mol. Med. Rep."},{"key":"B10","doi-asserted-by":"publisher","first-page":"258","DOI":"10.1016\/j.mam.2008.08.001","article-title":"The ADAM metalloproteinases","volume":"29","author":"Edwards","year":"2008","journal-title":"Mol. Asp. Med."},{"key":"B11","doi-asserted-by":"publisher","first-page":"859","DOI":"10.1093\/hmg\/ddi079","article-title":"A diastrophic dysplasia sulfate transporter (SLC26A2) mutant mouse: Morphological and biochemical characterization of the resulting chondrodysplasia phenotype","volume":"14","author":"Forlino","year":"2005","journal-title":"Hum. Mol. Genet."},{"key":"B12","doi-asserted-by":"publisher","first-page":"30469","DOI":"10.1074\/jbc.M213157200","article-title":"Catalytic activity of ADAM8, ADAM15, and MDC-L (ADAM28) on synthetic peptide substrates and in ectodomain cleavage of CD23","volume":"278","author":"Fourie","year":"2003","journal-title":"J. Biol. Chem."},{"key":"B13","doi-asserted-by":"publisher","first-page":"21214","DOI":"10.1074\/jbc.M112.347120","article-title":"ADAM15 protein amplifies focal adhesion kinase phosphorylation under genotoxic stress conditions","volume":"287","author":"Fried","year":"2012","journal-title":"J. Biol. Chem."},{"key":"B14","doi-asserted-by":"publisher","first-page":"100028","DOI":"10.1016\/j.jtauto.2019.100028","article-title":"Programmed death ligand 2 - a link between inflammation and bone loss in rheumatoid arthritis","volume":"3","author":"Greisen","year":"2020","journal-title":"J. Transl. Autoimmun."},{"key":"B15","doi-asserted-by":"publisher","first-page":"973","DOI":"10.1177\/002215540104900805","article-title":"SLC26A2 (diastrophic dysplasia sulfate transporter) is expressed in developing and mature cartilage but also in other tissues and cell types","volume":"49","author":"Haila","year":"2001","journal-title":"J. Histochem Cytochem"},{"key":"B16","first-page":"255","article-title":"Atelosteogenesis type II is caused by mutations in the diastrophic dysplasia sulfate-transporter gene (DTDST): Evidence for a phenotypic series involving three chondrodysplasias","volume":"58","author":"Hastbacka","year":"1996","journal-title":"Am. J. Hum. Genet."},{"key":"B17","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1111\/j.1749-6632.1996.tb56251.x","article-title":"Sulfate transport in chondrodysplasia","volume":"785","author":"Hastbacka","year":"1996","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"B18","doi-asserted-by":"publisher","first-page":"1487","DOI":"10.1074\/mcp.RA118.000608","article-title":"Click chemistry-mediated biotinylation reveals a function for the protease BACE1 in modulating the neuronal surface glycoproteome","volume":"17","author":"Herber","year":"2018","journal-title":"Mol. Cell Proteomics"},{"key":"B19","doi-asserted-by":"publisher","first-page":"5614","DOI":"10.1128\/mcb.23.16.5614-5624.2003","article-title":"Potential role for ADAM15 in pathological neovascularization in mice","volume":"23","author":"Horiuchi","year":"2003","journal-title":"Mol. Cell Biol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"3055","DOI":"10.1007\/s00018-019-03173-7","article-title":"Functions of 'A disintegrin and metalloproteases (ADAMs)' in the mammalian nervous system","volume":"76","author":"Hsia","year":"2019","journal-title":"Cell Mol. Life Sci."},{"key":"B21","doi-asserted-by":"publisher","first-page":"10732","DOI":"10.1073\/pnas.0404117101","article-title":"Vasorin, a transforming growth factor beta-binding protein expressed in vascular smooth muscle cells, modulates the arterial response to injury in vivo","volume":"101","author":"Ikeda","year":"2004","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B22","doi-asserted-by":"publisher","first-page":"1918","DOI":"10.1161\/ATVBAHA.120.314600","article-title":"ADAM (a disintegrin and metalloproteinase) 15 deficiency exacerbates ang II (angiotensin II)-Induced aortic remodeling leading to abdominal aortic aneurysm","volume":"40","author":"Jana","year":"2020","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"S117","DOI":"10.1016\/j.joca.2021.02.164","article-title":"Hyaluronan and proteoglycan link protein1 exhibits chondroregenerative ability in injury-induced osteoarthritic animals through ALK5-SMAD2 signaling in articular chondrocytes","volume":"29","author":"Jang","year":"2021","journal-title":"Osteoarthr. Cartil."},{"key":"B24","doi-asserted-by":"publisher","first-page":"741","DOI":"10.1515\/hsz-2012-0353","article-title":"Heparan sulfate: A key regulator of embryonic stem cell fate","volume":"394","author":"Kraushaar","year":"2013","journal-title":"Biol. Chem."},{"key":"B25","doi-asserted-by":"publisher","first-page":"261","DOI":"10.1038\/85330","article-title":"PD-L2 is a second ligand for PD-1 and inhibits T cell activation","volume":"2","author":"Latchman","year":"2001","journal-title":"Nat. Immunol."},{"key":"B26","doi-asserted-by":"publisher","first-page":"e99456","DOI":"10.15252\/embj.201899456","article-title":"Proteolytic ectodomain shedding of membrane proteins in mammals-hardware, concepts, and recent developments","volume":"37","author":"Lichtenthaler","year":"2018","journal-title":"EMBO J."},{"key":"B27","doi-asserted-by":"publisher","first-page":"6774","DOI":"10.1167\/iovs.14-14472","article-title":"Characterization of oxygen-induced retinopathy in mice carrying an inactivating point mutation in the catalytic site of ADAM15","volume":"55","author":"Maretzky","year":"2014","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"B28","doi-asserted-by":"publisher","first-page":"105","DOI":"10.1042\/BJ20082127","article-title":"Characterization of the catalytic activity of the membrane-anchored metalloproteinase ADAM15 in cell-based assays","volume":"420","author":"Maretzky","year":"2009","journal-title":"Biochem. J."},{"key":"B29","doi-asserted-by":"publisher","first-page":"12540","DOI":"10.1038\/s41598-019-49021-3","article-title":"ADAM15 mediates upregulation of Claudin-1 expression in breast cancer cells","volume":"9","author":"Mattern","year":"2019","journal-title":"Sci. Rep."},{"key":"B30","doi-asserted-by":"publisher","first-page":"846","DOI":"10.1038\/s41587-021-00860-4","article-title":"Ultra-fast proteomics with scanning SWATH","volume":"39","author":"Messner","year":"2021","journal-title":"Nat. Biotechnol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"18393","DOI":"10.1074\/jbc.M801329200","article-title":"The ectodomain shedding of E-cadherin by ADAM15 supports ErbB receptor activation","volume":"283","author":"Najy","year":"2008","journal-title":"J. Biol. Chem."},{"key":"B32","doi-asserted-by":"publisher","first-page":"663","DOI":"10.1021\/ac026117i","article-title":"Stop and go extraction tips for matrix-assisted laser desorption\/ionization, nanoelectrospray, and LC\/MS sample pretreatment in proteomics","volume":"75","author":"Rappsilber","year":"2003","journal-title":"Anal. Chem."},{"key":"B33","doi-asserted-by":"publisher","first-page":"47929","DOI":"10.1074\/jbc.M400129200","article-title":"Distinct ADAM metalloproteinases regulate G protein-coupled receptor-induced cell proliferation and survival","volume":"279","author":"Schafer","year":"2004","journal-title":"J. Biol. Chem."},{"key":"B34","doi-asserted-by":"publisher","first-page":"14002","DOI":"10.1038\/boneres.2014.2","article-title":"TGF-beta signaling and the development of osteoarthritis","volume":"2","author":"Shen","year":"2014","journal-title":"Bone Res."},{"key":"B35","doi-asserted-by":"publisher","first-page":"2856","DOI":"10.1038\/nprot.2006.468","article-title":"In-gel digestion for mass spectrometric characterization of proteins and proteomes","volume":"1","author":"Shevchenko","year":"2006","journal-title":"Nat. Protoc."},{"key":"B36","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1146\/annurev-bioeng-071516-044610","article-title":"Sulfation of glycosaminoglycans and its implications in human health and disorders","volume":"19","author":"Soares da Costa","year":"2017","journal-title":"Annu. Rev. Biomed. Eng."},{"key":"B37","doi-asserted-by":"publisher","first-page":"100","DOI":"10.1038\/ng0196-100","article-title":"Achondrogenesis type IB is caused by mutations in the diastrophic dysplasia sulphate transporter gene","volume":"12","author":"Superti-Furga","year":"1996","journal-title":"Nat. Genet."},{"key":"B38","doi-asserted-by":"publisher","first-page":"144","DOI":"10.1002\/(SICI)1096-8628(19960503)63:1<144::AID-AJMG25>3.0.CO;2-N","article-title":"A chondrodysplasia family produced by mutations in the diastrophic dysplasia sulfate transporter gene: Genotype\/phenotype correlations","volume":"63","author":"Superti-Furga","year":"1996","journal-title":"Am. J. Med. Genet."},{"key":"B39","doi-asserted-by":"publisher","first-page":"2017","DOI":"10.1016\/j.joca.2014.07.024","article-title":"Osteoarthritis year in review 2014: Genetics and genomics","volume":"22","author":"Tsezou","year":"2014","journal-title":"Osteoarthr. Cartil."},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.4238\/gmr.15027963","article-title":"Role of PPAR\u03b1 in down-regulating AGE-induced TGF-\u03b2 and MMP-9 expressions in chondrocytes","volume":"15","author":"Wang","year":"2016","journal-title":"Genet. Mol. Res."},{"key":"B41","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1038\/nmeth.1322","article-title":"Universal sample preparation method for proteome analysis","volume":"6","author":"Wisniewski","year":"2009","journal-title":"Nat. Methods"},{"key":"B42","doi-asserted-by":"publisher","first-page":"578","DOI":"10.3390\/membranes12060578","article-title":"Quantitative proteomics reveals that ADAM15 can have proteolytic-independent functions in the steady state","volume":"12","author":"Yang","year":"2022","journal-title":"Membr. (Basel)"},{"key":"B43","doi-asserted-by":"publisher","first-page":"100128","DOI":"10.1016\/j.ocarto.2020.100128","article-title":"Interleukin 13 (IL-13)-regulated expression of the chondroprotective metalloproteinase ADAM15 is reduced in aging cartilage","volume":"2","author":"Yang","year":"2020","journal-title":"Osteoarthr. Cartil. Open"},{"key":"B44","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1083\/jcb.153.1.35","article-title":"TGF-beta\/Smad3 signals repress chondrocyte hypertrophic differentiation and are required for maintaining articular cartilage","volume":"153","author":"Yang","year":"2001","journal-title":"J. Cell Biol."},{"key":"B45","doi-asserted-by":"publisher","first-page":"e1900276","DOI":"10.1002\/pmic.201900276","article-title":"Data-independent acquisition mass spectrometry-based proteomics and software tools: A glimpse in 2020","volume":"20","author":"Zhang","year":"2020","journal-title":"Proteomics"},{"key":"B46","doi-asserted-by":"publisher","first-page":"704","DOI":"10.1038\/nm.3143","article-title":"Inhibition of TGF-beta signaling in mesenchymal stem cells of subchondral bone attenuates osteoarthritis","volume":"19","author":"Zhen","year":"2013","journal-title":"Nat. Med."}],"container-title":["Frontiers in Molecular Biosciences"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmolb.2023.1162504\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,6,14]],"date-time":"2023-06-14T09:23:13Z","timestamp":1686734593000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmolb.2023.1162504\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,6,14]]},"references-count":46,"alternative-id":["10.3389\/fmolb.2023.1162504"],"URL":"https:\/\/doi.org\/10.3389\/fmolb.2023.1162504","relation":{},"ISSN":["2296-889X"],"issn-type":[{"value":"2296-889X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,6,14]]},"article-number":"1162504"}}