{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,29]],"date-time":"2026-10-29T17:11:50Z","timestamp":1793293910998,"version":"4.5.0"},"reference-count":25,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2016,2,22]],"date-time":"2016-02-22T00:00:00Z","timestamp":1456099200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"crossref","award":["K.O. 24592794"],"award-info":[{"award-number":["K.O. 24592794"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2016,3]]},"abstract":"<jats:p>Tissue\u2010nonspecific alkaline phosphatase (<jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content>) is a membrane glycoprotein with a proposed role in bone mineralization. Indeed, mutations in <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> have been identified in patients with hypophosphatasia (<jats:styled-content style=\"fixed-case\">HPP<\/jats:styled-content>), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum <jats:styled-content style=\"fixed-case\">ALP<\/jats:styled-content> activity. <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> has five potential N\u2010glycosylation sites at N140, N230, N271, N303 and N430 by standard nomenclature. A mutation at one of these sites, N430, was recently detected in a patient with infantile <jats:styled-content style=\"fixed-case\">HPP<\/jats:styled-content>. Using site\u2010directed mutagenesis, we demonstrated that <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> has five <jats:italic>N<\/jats:italic>\u2010glycans in transfected <jats:styled-content style=\"fixed-case\">COS<\/jats:styled-content>\u20101 cells and that individual single <jats:italic>N<\/jats:italic>\u2010glycan deletion mutants of <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> retain the dimeric structure required for <jats:styled-content style=\"fixed-case\">ALP<\/jats:styled-content> activity, excluding the possibility that any single <jats:italic>N<\/jats:italic>\u2010glycan plays a vital role in the structure and function of <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content>. However, we found that <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> (N430Q) and <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> (N430E) mutants, but not a <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> (N430D) mutant, failed to form dimers. The <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> (N430S) mutant linked to infantile <jats:styled-content style=\"fixed-case\">HPP<\/jats:styled-content> was glycosylation\u2010defective and unable to dimerise, similar to <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> (N430Q) and <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> (N430E) mutants; therefore, <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> (N430S) was established as a severe allele without strong <jats:styled-content style=\"fixed-case\">ALP<\/jats:styled-content> activity. By contrast to individual single <jats:italic>N<\/jats:italic>\u2010glycan deletion mutants, <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> devoid of all five <jats:italic>N<\/jats:italic>\u2010glycans was present to a much lesser extent than wild\u2010type <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> in transfected cells, possibly reflecting its instability. A comprehensive analysis of a series of multiple <jats:italic>N<\/jats:italic>\u2010glycan depletion mutants in <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> revealed that three <jats:italic>N<\/jats:italic>\u2010glycans on N230, N271 and N303 were the minimal requirement for the structure and function of <jats:styled-content style=\"fixed-case\">TNSALP<\/jats:styled-content> and a prerequisite for its stable expression in a cell.<\/jats:p>","DOI":"10.1111\/febs.13663","type":"journal-article","created":{"date-parts":[[2016,1,21]],"date-time":"2016-01-21T17:19:45Z","timestamp":1453396785000},"page":"1168-1179","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Glycosylation\u2010deficient mutations in tissue\u2010nonspecific alkaline phosphatase impair its structure and function and are linked to infantile hypophosphatasia"],"prefix":"10.1111","volume":"283","author":[{"given":"Keiichi","family":"Komaru","sequence":"first","affiliation":[{"name":"Kitasato Junior College of Health and Hygienic Sciences Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasuhito","family":"Satou","sequence":"additional","affiliation":[{"name":"Division of Oral Biochemistry Niigata University Graduate School of Medical and Dental Sciences Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiba A.","family":"Al\u2010Shawafi","sequence":"additional","affiliation":[{"name":"Division of Oral Biochemistry Niigata University Graduate School of Medical and Dental Sciences Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natsuko","family":"Numa\u2010Kinjoh","sequence":"additional","affiliation":[{"name":"Division of Oral Biochemistry Niigata University Graduate School of Medical and Dental Sciences 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