{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T01:13:27Z","timestamp":1789521207452,"version":"build-2803163510"},"reference-count":29,"publisher":"Portland Press Ltd.","issue":"1","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2014,10,1]]},"abstract":"<jats:p>OPN (osteopontin) is a multiphosphorylated extracellular glycoprotein, which has important roles in bone remodelling, inflammation and cancer metastasis. OPN regulates cell spreading and adhesion primarily through its association with several integrins such as \u03b1v\u03b23, and its phosphorylation affects these processes. However, the mechanism by which OPN O-glycosylation affects these processes is not completely understood. In the present study, we demonstrated that OPN O-glycosylation self-regulates its biological activities and also affects its phosphorylation status. We prepared two recombinant OPNs, WT (wild-type)-OPN and mutant OPN (\u0394O-OPN), which lacks five O-glycosylation sites at a threonine\/proline-rich region. O-glycan defects in OPN increased its phosphorylation level, as observed by dephosphorylation assays. Moreover, compared with WT-OPN, \u0394O-OPN exhibited enhanced cell spreading and adhesion activities and decreased associations with \u03b21 integrins. This suggested that defects in O-glycans in OPN altered these activities, and that \u03b21 integrins have a less important role in adhesion to \u0394O-OPN. The cell-adhesion activity of dephosphorylated \u0394O-OPN was higher than the cell-adhesion activities of \u0394O-OPN and dephosphorylated WT-OPN. This suggested that some of the phosphorylation in \u0394O-OPN caused by O-glycan defects and O-glycans of OPN suppressed the OPN cell-adhesion activity. Thus functional activities of OPN can be determined by the combined glycosylation and phosphorylation statuses and not by either status alone.<\/jats:p>","DOI":"10.1042\/bj20140060","type":"journal-article","created":{"date-parts":[[2014,7,16]],"date-time":"2014-07-16T12:36:02Z","timestamp":1405514162000},"page":"93-102","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":52,"title":["Osteopontin <i>O<\/i>-glycosylation contributes to its phosphorylation and cell-adhesion properties"],"prefix":"10.1042","volume":"463","author":[{"given":"Yoshinobu","family":"Kariya","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Fukushima Medical University School of Medicine, 1 Hikarigaoka, Fukushima City, Fukushima 960-1295, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mayumi","family":"Kanno","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Fukushima Medical University School of Medicine, 1 Hikarigaoka, Fukushima City, Fukushima 960-1295, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kana","family":"Matsumoto-Morita","sequence":"additional","affiliation":[{"name":"Structural Glycobiology Team, Systems Glycobiology Research Group, RIKEN-Max Planck Joint Research Center, RIKEN Global Research Cluster, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Midori","family":"Konno","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Fukushima Medical University School of Medicine, 1 Hikarigaoka, Fukushima City, Fukushima 960-1295, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshiki","family":"Yamaguchi","sequence":"additional","affiliation":[{"name":"Structural Glycobiology Team, Systems Glycobiology Research Group, RIKEN-Max Planck Joint Research Center, RIKEN Global Research Cluster, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasuhiro","family":"Hashimoto","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Fukushima Medical University School of Medicine, 1 Hikarigaoka, Fukushima City, Fukushima 960-1295, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2014,9,8]]},"reference":[{"key":"2021112216074933700_B1","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1177\/10454411000110030101","article-title":"Osteopontin","volume":"11","author":"Sodek","year":"2000","journal-title":"Crit. Rev. Oral Biol. Med."},{"key":"2021112216074933700_B2","doi-asserted-by":"publisher","first-page":"912","DOI":"10.1002\/jcb.21558","article-title":"Control of osteopontin signaling and function by post-translational phosphorylation and protein folding","volume":"102","author":"Kazanecki","year":"2007","journal-title":"J. Cell Biochem."},{"key":"2021112216074933700_B3","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1042\/bj2320715","article-title":"Isolation and characterization of two sialoproteins present only in bone calcified matrix","volume":"232","author":"Franzen","year":"1985","journal-title":"Biochem. J."},{"key":"2021112216074933700_B4","doi-asserted-by":"publisher","first-page":"285","DOI":"10.1042\/BJ20050341","article-title":"Post-translationally modified residues of native human osteopontin are located in clusters identification of 36 phosphorylation and five O-glycosylation sites and their biological implications","volume":"390","author":"Christensen","year":"2005","journal-title":"Biochem. J."},{"key":"2021112216074933700_B5","doi-asserted-by":"publisher","first-page":"2040","DOI":"10.1002\/pro.5560041009","article-title":"Posttranslational modifications of bovine osteopontin: identification of twenty-eight phosphorylation and three O-glycosylation sites","volume":"4","author":"Sorensen","year":"1995","journal-title":"Protein Sci."},{"key":"2021112216074933700_B6","doi-asserted-by":"crossref","first-page":"2900","DOI":"10.1016\/S0021-9258(18)61592-3","article-title":"Isolation, characterization, and biosynthesis of a phosphorylated glycoprotein from rat bone","volume":"262","author":"Prince","year":"1987","journal-title":"J. Biol. Chem."},{"key":"2021112216074933700_B7","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1042\/BJ20071021","article-title":"Post-translational modification and proteolytic processing of urinary osteopontin","volume":"411","author":"Christensen","year":"2008","journal-title":"Biochem. J."},{"key":"2021112216074933700_B8","doi-asserted-by":"publisher","first-page":"3788","DOI":"10.1074\/jbc.M111.277996","article-title":"C-terminal modification of osteopontin inhibits interaction with the alphaVbeta3-integrin","volume":"287","author":"Christensen","year":"2012","journal-title":"J. Biol. Chem."},{"key":"2021112216074933700_B9","doi-asserted-by":"crossref","first-page":"14853","DOI":"10.1016\/S0021-9258(17)36541-9","article-title":"Dephosphorylation of osteopontin and bone sialoprotein by osteoclastic tartrate-resistant acid phosphatase. Modulation of osteoclast adhesion in\u00a0vitro","volume":"269","author":"Ek-Rylander","year":"1994","journal-title":"J. Biol. Chem."},{"key":"2021112216074933700_B10","doi-asserted-by":"crossref","first-page":"18202","DOI":"10.1016\/S0021-9258(19)84697-5","article-title":"Normal rat kidney cells secrete both phosphorylated and nonphosphorylated forms of osteopontin showing different physiological properties","volume":"264","author":"Nemir","year":"1989","journal-title":"J. Biol. Chem."},{"key":"2021112216074933700_B11","doi-asserted-by":"publisher","first-page":"860","DOI":"10.1126\/science.287.5454.860","article-title":"Eta-1 (osteopontin): an early component of type-1 (cell-mediated) immunity","volume":"287","author":"Ashkar","year":"2000","journal-title":"Science"},{"key":"2021112216074933700_B12","doi-asserted-by":"publisher","first-page":"5729","DOI":"10.1021\/bi961687w","article-title":"Altered sialylation of osteopontin prevents its receptor-mediated binding on the surface of oncogenically transformed tsB77 cells","volume":"36","author":"Shanmugam","year":"1997","journal-title":"Biochemistry"},{"key":"2021112216074933700_B13","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1007\/s10549-004-3137-3","article-title":"Stable expression of sialyl-Tn antigen in T47-D cells induces a decrease of cell adhesion and an increase of cell migration","volume":"90","author":"Julien","year":"2005","journal-title":"Breast Cancer Res. Treat."},{"key":"2021112216074933700_B14","doi-asserted-by":"publisher","first-page":"5059","DOI":"10.1242\/jcs.01350","article-title":"Expression of sialyl-Tn epitopes on beta1 integrin alters epithelial cell phenotype, proliferation and haptotaxis","volume":"117","author":"Clement","year":"2004","journal-title":"J. Cell Sci."},{"key":"2021112216074933700_B15","doi-asserted-by":"publisher","first-page":"19463","DOI":"10.1074\/jbc.M703055200","article-title":"Cell type-specific post-translational modifications of mouse osteopontin are associated with different adhesive properties","volume":"282","author":"Christensen","year":"2007","journal-title":"J. Biol. Chem."},{"key":"2021112216074933700_B16","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1038\/nrc1251","article-title":"Mucins in cancer: protection and control of the cell surface","volume":"4","author":"Hollingsworth","year":"2004","journal-title":"Nat. Rev. Cancer."},{"key":"2021112216074933700_B17","doi-asserted-by":"publisher","first-page":"204","DOI":"10.1186\/bcr2577","article-title":"Tumour-associated carbohydrate antigens in breast cancer","volume":"12","author":"Cazet","year":"2010","journal-title":"Breast Cancer Res."},{"key":"2021112216074933700_B18","doi-asserted-by":"publisher","first-page":"1746","DOI":"10.1038\/sj.bjc.6605083","article-title":"Sialyl-Tn vaccine induces antibody-mediated tumour protection in a relevant murine model","volume":"100","author":"Julien","year":"2009","journal-title":"Br. J. Cancer."},{"key":"2021112216074933700_B19","doi-asserted-by":"publisher","first-page":"607","DOI":"10.1093\/oxfordjournals.jbchem.a003263","article-title":"Efficient expression system of human recombinant laminin-5","volume":"132","author":"Kariya","year":"2002","journal-title":"J. Biochem."},{"key":"2021112216074933700_B20","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":"2021112216074933700_B21","doi-asserted-by":"publisher","first-page":"5354","DOI":"10.1021\/pr200611n","article-title":"Universal and confident phosphorylation site localization using phosphoRS","volume":"10","author":"Taus","year":"2011","journal-title":"J. Proteome Res."},{"key":"2021112216074933700_B22","doi-asserted-by":"publisher","first-page":"265","DOI":"10.1083\/jcb.201308136","article-title":"Talin-bound NPLY motif recruits integrin-signaling adapters to regulate cell spreading and mechanosensing","volume":"205","author":"Pinon","year":"2014","journal-title":"J. Cell Biol."},{"key":"2021112216074933700_B23","doi-asserted-by":"publisher","first-page":"e27084","DOI":"10.1371\/journal.pone.0027084","article-title":"N-glycosylation of \u03b24 integrin controls the adhesion and motility of keratinocytes","volume":"6","author":"Kariya","year":"2011","journal-title":"PLoS ONE"},{"key":"2021112216074933700_B24","doi-asserted-by":"publisher","first-page":"22200","DOI":"10.1074\/jbc.M102150200","article-title":"Structural elucidation of the N- and O-glycans of human apolipoprotein(a): role of O-glycans in conferring protease resistance","volume":"276","author":"Garner","year":"2001","journal-title":"J. Biol. Chem."},{"key":"2021112216074933700_B25","first-page":"8278","article-title":"Monoclonal antibodies with defined recognition sequences in the stem region of CD44: detection of differential glycosylation of CD44 between tumor and stromal cells in tissue","volume":"63","author":"Matsuki","year":"2003","journal-title":"Cancer Res."},{"key":"2021112216074933700_B26","doi-asserted-by":"crossref","first-page":"18696","DOI":"10.1016\/S0021-9258(17)44808-3","article-title":"Physiological properties and differential glycosylation of phosphorylated and nonphosphorylated forms of osteopontin secreted by normal rat kidney cells","volume":"265","author":"Singh","year":"1990","journal-title":"J. Biol. Chem."},{"key":"2021112216074933700_B27","doi-asserted-by":"publisher","first-page":"1150","DOI":"10.1126\/science.1217817","article-title":"Secreted kinase phosphorylates extracellular proteins that regulate biomineralization","volume":"336","author":"Tagliabracci","year":"2012","journal-title":"Science"},{"key":"2021112216074933700_B28","doi-asserted-by":"publisher","first-page":"1017","DOI":"10.1038\/nature05815","article-title":"Cycling of O-linked beta-N-acetylglucosamine on nucleocytoplasmic proteins","volume":"446","author":"Hart","year":"2007","journal-title":"Nature"},{"key":"2021112216074933700_B29","doi-asserted-by":"publisher","first-page":"599","DOI":"10.1038\/sj.embor.7400705","article-title":"Mucin-type O-glycans in human colon and breast cancer glycodynamics and functions","volume":"7","author":"Brockhausen","year":"2006","journal-title":"EMBO Rep."}],"container-title":["Biochemical Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/463\/1\/93\/681340\/bj4630093.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/463\/1\/93\/681340\/bj4630093.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,22]],"date-time":"2021-11-22T22:57:24Z","timestamp":1637621844000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemj\/article\/463\/1\/93\/47950\/Osteopontin-O-glycosylation-contributes-to-its"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,9,8]]},"references-count":29,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2014,10,1]]}},"URL":"https:\/\/doi.org\/10.1042\/bj20140060","relation":{},"ISSN":["0264-6021","1470-8728"],"issn-type":[{"value":"0264-6021","type":"print"},{"value":"1470-8728","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,9,8]]}}}