{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,5]],"date-time":"2025-12-05T12:06:20Z","timestamp":1764936380535},"reference-count":31,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2005,3,3]],"date-time":"2005-03-03T00:00:00Z","timestamp":1109808000000},"content-version":"vor","delay-in-days":3471,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["European Journal of Biochemistry"],"published-print":{"date-parts":[[1995,9]]},"abstract":"<jats:p>The human \u03b2\u2010trace protein has been cloned and has been expressed for the first time in a mammalian host cell line. Stable BHK\u201021 cell lines exhibiting altered terminal sialylation properties were constructed by cotransfection of cells with the plasmids pMT\u2010\u03b2TP or pAB3\u20101 which contain the cDNAs encoding the human secretory glycoproteins \u03b2\u2010trace protein or antithrombin III and pABSial containing the human Golgi enzyme CMP\u2010NeuAc:Gal(\u03b21\u20104)GlcNAc\u2010R \u03b12,6\u2010sialyltransferase (ST6N) gene. The \u03b2\u2010trace protein was purified by immunoaffinity chromatography and N\u2010linked oligosaccharides were subjected to carbohydrate structural analysis. The enzymically liberated oligosaccharides were found to consist of 90% of diantennary chains as is the case for natural \u03b2\u2010trace protein from human cerebrospinal fluid. About 90% of the total oligosaccharides were recovered in the monosialo and disialo fractions in a ratio of 1:5. The monosialylated oligosaccharides of \u03b2\u2010trace protein coexpressed with human ST6N were found to contain NeuAc in \u03b12,6\u2010or \u03b12,3\u2010linkage in the same ratio. From <jats:sup>1<\/jats:sup>H\u2010NMR analysis as well as calculations of peak areas obtained by HPLC, 60% of the molecules of the disialo fraction were found to contain NeuAc in both \u03b12,3\u2010and \u03b12,6\u2010linkage to Gal\u03b2(1\u20104)GlcNAc\u2010R, whereas 40% of the molecules of this fraction contained NeuAc in only \u03b12,3\u2010linkage to Gal(\u03b21\u20104)GlcNAc\u2010R. The \u03b12,6\u2010linked NeuAc was shown to be attached preferentially to the Gal(\u03b21\u20104)GlcNAc(\u03b21\u20102)Man(\u03b11\u20103) branch of the diantennary structure. Therefore the <jats:italic>in vivo<\/jats:italic> specificity of the newly introduced recombinant human ST6N observed in this study supports the previously reported <jats:italic>in vitro<\/jats:italic> branch specificity of the bovine colostrum ST6N activity. Furthermore, these studies demonstrate the suitability of genetically engineered mammalian host cell lines with novel glycosylation properties for the production of human\u2010type glycosylated secretory recombinant polypeptides.<\/jats:p>","DOI":"10.1111\/j.1432-1033.1995.718zz.x","type":"journal-article","created":{"date-parts":[[2005,3,4]],"date-time":"2005-03-04T12:51:36Z","timestamp":1109940696000},"page":"718-725","source":"Crossref","is-referenced-by-count":59,"title":["Construction of Stable BHK\u201021 Cells Coexpressing Human Secretory Glycoproteins and Human Gal(\u03b21\u20104)GlcNAc\u2010R \u03b12,6\u2010Sialyltransferase"],"prefix":"10.1111","volume":"232","author":[{"given":"Eckart","family":"Grabenhorst","sequence":"first","affiliation":[]},{"given":"Andrea","family":"Hoffman","sequence":"additional","affiliation":[]},{"given":"Manfred","family":"Nimtz","sequence":"additional","affiliation":[]},{"given":"Gerd","family":"Zettlmeissl","sequence":"additional","affiliation":[]},{"given":"Harald S.","family":"Conradt","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2008,6,28]]},"reference":[{"key":"e_1_2_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)47838-6"},{"key":"e_1_2_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)45317-3"},{"key":"e_1_2_3_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)80078-3"},{"key":"e_1_2_3_5_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1993.tb17732.x"},{"key":"e_1_2_3_6_2","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1042\/bj2940025","volume":"294","author":"Meier W.","year":"1993","journal-title":"Biochem. 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