{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T19:01:00Z","timestamp":1791313260136,"version":"4.3.1"},"reference-count":42,"publisher":"American Chemical Society (ACS)","issue":"8","license":[{"start":{"date-parts":[[2015,6,23]],"date-time":"2015-06-23T00:00:00Z","timestamp":1435017600000},"content-version":"vor","delay-in-days":365,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2014,8,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Collagens are the most abundant proteins in animals and are involved in many physiological\/pathological events. Although various methods have been used to quantify collagen and its post-translational modifications (PTMs) over the years, it is still difficult to accurately quantify type-specific collagen and minor collagen PTMs. We report a novel quantitative method targeting collagen using stable isotope-labeled collagen named \u201cSI-collagen\u201d, which was labeled with isotopically heavy lysine, arginine, and proline in fibroblasts culture. We prepared highly labeled and purified SI-collagen for use as an internal standard in mass spectrometric analysis, particularly for a new approach using amino acid hydrolysis. Our method enabled accurate collagen analyses, including quantification of (1) type-specific collagen (types I and III in this paper), (2) total collagen, and (3) collagen PTMs by LC\u2013MS with high sensitivity. SI-collagen is also applicable to other diverse analyses of collagen and can be a powerful tool for various studies, such as detailed investigation of collagen-related disorders.<\/jats:p>","DOI":"10.1021\/pr500213a","type":"journal-article","created":{"date-parts":[[2014,6,23]],"date-time":"2014-06-23T05:50:46Z","timestamp":1403502646000},"page":"3671-3678","source":"Crossref","is-referenced-by-count":49,"title":["Stable Isotope-Labeled Collagen:\nA Novel and Versatile\nTool for Quantitative Collagen Analyses Using Mass Spectrometry"],"prefix":"10.1021","volume":"13","author":[{"given":"Yuki","family":"Taga","sequence":"first","affiliation":[{"name":"Nippi Research Institute of Biomatrix , 520-11 Kuwabara , , ,","place":["Toride, Ibaraki, Japan, 302-0017"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masashi","family":"Kusubata","sequence":"additional","affiliation":[{"name":"Nippi Research Institute of Biomatrix , 520-11 Kuwabara , , ,","place":["Toride, Ibaraki, Japan, 302-0017"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kiyoko","family":"Ogawa-Goto","sequence":"additional","affiliation":[{"name":"Nippi Research Institute of Biomatrix , 520-11 Kuwabara , , ,","place":["Toride, Ibaraki, Japan, 302-0017"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shunji","family":"Hattori","sequence":"additional","affiliation":[{"name":"Nippi Research Institute of Biomatrix , 520-11 Kuwabara , , ,","place":["Toride, Ibaraki, Japan, 302-0017"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2014,6,23]]},"reference":[{"issue":"6","key":"2026090309345321900_cit1","doi-asserted-by":"crossref","first-page":"1498","DOI":"10.1172\/JCI108420","article-title":"Collagen polymorphism in idiopathic\nchronic pulmonary fibrosis","volume":"57","author":"Seyer","year":"1976","journal-title":"J. Clin. Invest."},{"issue":"2","key":"2026090309345321900_cit2","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1172\/JCI108634","article-title":"Collagen polymorphism in normal and\ncirrhotic human liver","volume":"59","author":"Seyer","year":"1977","journal-title":"J. Clin. Invest."},{"issue":"11","key":"2026090309345321900_cit3","first-page":"L709","article-title":"Always cleave\nup your mess: targeting\ncollagen degradation to treat tissue fibrosis","volume":"304","author":"McKleroy","year":"2013","journal-title":"Am. J. Physiol.: Lung Cell. Mol. Physiol."},{"issue":"9","key":"2026090309345321900_cit4","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1038\/nrendo.2011.81","article-title":"New perspectives\non osteogenesis imperfecta","volume":"7","author":"Forlino","year":"2011","journal-title":"Nat. Rev. Endocrinol"},{"key":"2026090309345321900_cit5","doi-asserted-by":"crossref","DOI":"10.1093\/oso\/9780199632213.001.0001","volume-title":"Extracellular Matrix:\nA Practical Approach","author":"Haralson","year":"1995"},{"key":"2026090309345321900_cit6","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/0076-6879(82)82071-5","article-title":"Hydroxylysine\nglycosides","volume":"82","author":"Butler","year":"1982","journal-title":"Methods Enzymol."},{"key":"2026090309345321900_cit7","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1016\/0076-6879(82)82074-0","article-title":"Determination\nof 3- and 4-hydroxyproline","volume":"82","author":"Berg","year":"1982","journal-title":"Methods Enzymol."},{"key":"2026090309345321900_cit8","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/B978-0-12-363709-3.50012-7","article-title":"Localization of collagen types in tissues","volume":"9","author":"von der Mark","year":"1981","journal-title":"Int. Rev. Connect. Tissue Res."},{"issue":"3","key":"2026090309345321900_cit9","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1007\/s00726-005-0300-z","article-title":"Amino acid analysis by hydrophilic\ninteraction chromatography coupled on-line to electrospray ionization\nmass spectrometry","volume":"30","author":"Langrock","year":"2006","journal-title":"Amino Acids"},{"issue":"20","key":"2026090309345321900_cit10","doi-asserted-by":"crossref","first-page":"3483","DOI":"10.1002\/jssc.200800351","article-title":"Identification of collagen types\nin tissues using HPLC-MS\/MS","volume":"31","author":"Pataridis","year":"2008","journal-title":"J. Sep Sci."},{"issue":"4","key":"2026090309345321900_cit11","doi-asserted-by":"crossref","first-page":"2580","DOI":"10.1074\/jbc.M109.068726","article-title":"Location of 3-hydroxyproline\nresidues in collagen types\nI, II, III, and V\/XI implies a role in fibril supramolecular assembly","volume":"285","author":"Weis","year":"2010","journal-title":"J. Biol. Chem."},{"issue":"26","key":"2026090309345321900_cit12","doi-asserted-by":"crossref","first-page":"22253","DOI":"10.1074\/jbc.M111.333336","article-title":"Mutation\nin cyclophilin B that causes hyperelastosis cutis in American Quarter\nHorse does not affect peptidylprolyl cis-trans isomerase activity\nbut shows altered cyclophilin B-protein interactions and affects collagen\nfolding","volume":"287","author":"Ishikawa","year":"2012","journal-title":"J. Biol. Chem."},{"issue":"27","key":"2026090309345321900_cit13","doi-asserted-by":"crossref","first-page":"22998","DOI":"10.1074\/jbc.M112.343954","article-title":"Lysyl hydroxylase 3-mediated glucosylation\nin type I collagen: molecular loci and biological significance","volume":"287","author":"Sricholpech","year":"2012","journal-title":"J. Biol. Chem."},{"issue":"48","key":"2026090309345321900_cit14","doi-asserted-by":"crossref","first-page":"40598","DOI":"10.1074\/jbc.M112.406850","article-title":"Comprehensive mass spectrometric\nmapping of the hydroxylated amino acid residues of the alpha1(V) collagen\nchain","volume":"287","author":"Yang","year":"2012","journal-title":"J. Biol. Chem."},{"issue":"5","key":"2026090309345321900_cit15","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1074\/mcp.M200025-MCP200","article-title":"Stable isotope\nlabeling\nby amino acids in cell culture, SILAC, as a simple and accurate approach\nto expression proteomics","volume":"1","author":"Ong","year":"2002","journal-title":"Mol. Cell. Proteomics"},{"issue":"5","key":"2026090309345321900_cit16","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1038\/nbt1086","article-title":"Quantitative\nmouse brain proteomics\nusing culture-derived isotope tags as internal standards","volume":"23","author":"Ishihama","year":"2005","journal-title":"Nat. Biotechnol."},{"issue":"5","key":"2026090309345321900_cit17","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1038\/nmeth.1446","article-title":"Super-SILAC mix for\nquantitative proteomics of human tumor tissue","volume":"7","author":"Geiger","year":"2010","journal-title":"Nat. Methods"},{"issue":"3","key":"2026090309345321900_cit18","doi-asserted-by":"crossref","first-page":"1118","DOI":"10.1021\/pr7007175","article-title":"Absolute SILAC for\naccurate quantitation of proteins in complex mixtures down to the\nattomole level","volume":"7","author":"Hanke","year":"2008","journal-title":"J. Proteome Res."},{"issue":"6","key":"2026090309345321900_cit19","doi-asserted-by":"crossref","first-page":"1584","DOI":"10.1084\/jem.147.6.1584","article-title":"Affinity of fibronectin to collagens\nof different genetic types and to fibrinogen","volume":"147","author":"Engvall","year":"1978","journal-title":"J. Exp. Med."},{"issue":"2","key":"2026090309345321900_cit20","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1016\/0005-2795(73)90271-7","article-title":"The influence of dietary\ncalcium deficiency and parathyroidectomy on bone collagen structure","volume":"328","author":"Barnes","year":"1973","journal-title":"Biochim. Biophys. Acta"},{"issue":"4","key":"2026090309345321900_cit21","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1093\/oxfordjournals.jbchem.a131324","article-title":"Comparative study of carbohydrate-protein\ncomplexes. II. Determination of hydroxylysine and its glycosides in\nhuman skin and scar collagens by an improved method","volume":"80","author":"Isemura","year":"1976","journal-title":"J. Biochem."},{"issue":"39","key":"2026090309345321900_cit22","doi-asserted-by":"crossref","first-page":"29941","DOI":"10.1074\/jbc.M109.094607","article-title":"Enhancement\nof procollagen biosynthesis by p180 through augmented ribosome association\non the endoplasmic reticulum in response to stimulated secretion","volume":"285","author":"Ueno","year":"2010","journal-title":"J. Biol. Chem."},{"key":"2026090309345321900_cit23","doi-asserted-by":"crossref","first-page":"1005","DOI":"10.1146\/annurev.bi.49.070180.005041","article-title":"Metabolism of proline and the hydroxyprolines","volume":"49","author":"Adams","year":"1980","journal-title":"Annu. Rev. Biochem."},{"issue":"9","key":"2026090309345321900_cit24","doi-asserted-by":"crossref","first-page":"1587","DOI":"10.1074\/mcp.M800113-MCP200","article-title":"Prevention of amino acid conversion in SILAC experiments\nwith embryonic stem cells","volume":"7","author":"Bendall","year":"2008","journal-title":"Mol. Cell. Proteomics"},{"issue":"1","key":"2026090309345321900_cit25","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.ab.2011.01.011","article-title":"Preventing\narginine-to-proline conversion in a cell-line-independent manner during\ncell cultivation under stable isotope labeling by amino acids in cell\nculture (SILAC) conditions","volume":"412","author":"Lossner","year":"2011","journal-title":"Anal. Biochem."},{"issue":"10","key":"2026090309345321900_cit26","doi-asserted-by":"crossref","first-page":"3944","DOI":"10.1016\/S0021-9258(18)97133-4","article-title":"End product control of amino acid\nsynthesis by cultured human cells","volume":"240","author":"Eagle","year":"1965","journal-title":"J. Biol.\nChem."},{"issue":"Pt A","key":"2026090309345321900_cit27","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/0076-6879(82)82058-2","article-title":"Collagen: an overview","volume":"82","author":"Miller","year":"1982","journal-title":"Methods Enzymol."},{"issue":"20","key":"2026090309345321900_cit28","doi-asserted-by":"crossref","first-page":"4813","DOI":"10.1016\/S0021-9258(18)99529-3","article-title":"The structure\nof the disaccharide unit of the renal glomerular basement membrane","volume":"242","author":"Spiro","year":"1967","journal-title":"J. Biol. Chem."},{"issue":"46","key":"2026090309345321900_cit29","doi-asserted-by":"crossref","first-page":"27711","DOI":"10.1074\/jbc.270.46.27711","article-title":"Use of a new rat chondrosarcoma cell\nline to delineate\na 119-base pair chondrocyte-specific enhancer element and to define\nactive promoter segments in the mouse pro-alpha 1(II) collagen gene","volume":"270","author":"Mukhopadhyay","year":"1995","journal-title":"J. Biol. Chem."},{"issue":"1","key":"2026090309345321900_cit30","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1177\/39.1.1983874","article-title":"Human bone contains\ntype III collagen,\ntype VI collagen, and fibrillin: type III collagen is present on specific\nfibers that may mediate attachment of tendons, ligaments, and periosteum\nto calcified bone cortex","volume":"39","author":"Keene","year":"1991","journal-title":"J. Histochem. Cytochem."},{"issue":"10","key":"2026090309345321900_cit31","doi-asserted-by":"crossref","first-page":"7732","DOI":"10.1074\/jbc.C110.195768","article-title":"A novel 3-hydroxyproline\n(3Hyp)-rich motif marks the triple-helical C terminus of tendon type\nI collagen","volume":"286","author":"Eyre","year":"2011","journal-title":"J. Biol. Chem."},{"issue":"4","key":"2026090309345321900_cit32","doi-asserted-by":"crossref","first-page":"602","DOI":"10.1016\/S0021-9258(18)94398-X","article-title":"Characterization\nand quantitative determination of the hydroxylysine-linked carbohydrate\nunits of several collagens","volume":"244","author":"Spiro","year":"1969","journal-title":"J. Biol. Chem."},{"issue":"2","key":"2026090309345321900_cit33","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1016\/0005-2795(73)90130-X","article-title":"Increased lysine hydroxylation in\nrat bone and tendon collagen and localization of the additional residues","volume":"310","author":"Stoltz","year":"1973","journal-title":"Biochim. Biophys. Acta"},{"issue":"5","key":"2026090309345321900_cit34","first-page":"1009","article-title":"Collagen polymorphism in extracellular matrix of human\nosteosarcoma","volume":"69","author":"Shapiro","year":"1982","journal-title":"J. Natl. Cancer Inst."},{"issue":"7","key":"2026090309345321900_cit35","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1007\/BF01212948","article-title":"Composition\nand posttranslational modification of individual\ncollagen chains from osteosarcomas and osteofibrous dysplasias","volume":"121","author":"Lehmann","year":"1995","journal-title":"J. Cancer Res. Clin. Oncol."},{"issue":"2","key":"2026090309345321900_cit36","first-page":"113","article-title":"Increased\ntype-III\/type-I collagen\nratios in diabetic human conjunctival biopsies","volume":"4","author":"Kern","year":"1986","journal-title":"Clin. Physiol. Biochem."},{"issue":"12","key":"2026090309345321900_cit37","doi-asserted-by":"crossref","first-page":"1695","DOI":"10.2337\/diab.37.12.1695","article-title":"Phenotypic expression of collagen\ntypes in mesangial matrix of diabetic and nondiabetic rats","volume":"37","author":"Abrass","year":"1988","journal-title":"Diabetes"},{"issue":"2","key":"2026090309345321900_cit38","doi-asserted-by":"crossref","first-page":"119","DOI":"10.3109\/03008209309014239","article-title":"Biochemical changes\nin the collagen of human osteoporotic bone matrix","volume":"29","author":"Bailey","year":"1993","journal-title":"Connect Tissue Res."},{"issue":"6","key":"2026090309345321900_cit39","doi-asserted-by":"crossref","first-page":"M111.010397","DOI":"10.1074\/mcp.M111.010397","article-title":"Development of a novel\nmethod for analyzing collagen O-glycosylations by hydrazide chemistry","volume":"11","author":"Taga","year":"2012","journal-title":"Mol. Cell. Proteomics"},{"issue":"5","key":"2026090309345321900_cit40","doi-asserted-by":"crossref","first-page":"2225","DOI":"10.1021\/pr400079d","article-title":"Site-specific Quantitative\nAnalysis of Overglycosylation of Collagen in Osteogenesis Imperfecta\nUsing Hydrazide Chemistry and SILAC","volume":"12","author":"Taga","year":"2013","journal-title":"J. Proteome\nRes."},{"issue":"2","key":"2026090309345321900_cit41","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1016\/j.talanta.2011.01.050","article-title":"A systematic approach\nto assess amino acid conversions in SILAC experiments","volume":"84","author":"Marcilla","year":"2011","journal-title":"Talanta"},{"issue":"37","key":"2026090309345321900_cit42","doi-asserted-by":"crossref","first-page":"27037","DOI":"10.1074\/jbc.M701088200","article-title":"Extracellular collagenases\nand the endocytic receptor, urokinase plasminogen activator receptor-associated\nprotein\/Endo180, cooperate in fibroblast-mediated collagen degradation","volume":"282","author":"Madsen","year":"2007","journal-title":"J. Biol. Chem."}],"container-title":["Journal of Proteome Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/pr500213a","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jprobs\/article-pdf\/13\/8\/3671\/15493440\/pr500213a.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jprobs\/article-pdf\/13\/8\/3671\/15493440\/pr500213a.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T13:35:01Z","timestamp":1788442501000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jprobs\/article\/13\/8\/3671\/1463428\/Stable-Isotope-Labeled-Collagen-A-Novel-and"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,6,23]]},"references-count":42,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2014,6,23]]},"published-print":{"date-parts":[[2014,8,1]]}},"URL":"https:\/\/doi.org\/10.1021\/pr500213a","relation":{},"ISSN":["1535-3893","1535-3907"],"issn-type":[{"value":"1535-3893","type":"print"},{"value":"1535-3907","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,6,23]]}}}