caa a22 4500 469114959 CHVBK 20180323133122.0 cr unu---uuuuu 170328e19920201xx s 000 0 eng 10.1007/BF00731176 doi (NATIONALLICENCE)springer-10.1007/BF00731176 Expression, glycosylation and secretion of yeast acid phosphatase in hamster BHK cells [Elektronische Daten] [Rajko Reljic, Slobodan Barbaric, Blanka Ries, Roger Buxton, R. Hughes] The gene PHO5 coding for one of the repressible acid phosphatases of the yeastSaccharomyces cerevisiae has been expressed at high efficiency in the baby hamster kidney (BHK) cell line. The expression vector was constructed from PHO5 driven by the human β-actin promoter and was transfected into BHK cells by the calcium phosphate method. The recombinant APase (r-APase) which was secreted in active form from the cells was estimated by SDS/polyacrylamide gel electrophoresis to have molecular massM r=62000, indicating substitution of the polypeptide moiety by 2-3 asparagine-linked glycans. Analysis by sequential lectin affinity chromatography of glycopeptides obtained from r-APase with Pronase showed that the glycans are predominantly of the 2.2.4 triantennary and tetraantennary complex-type. These data suggest that the extensive glycosylation of yeast APase, which contains eight polymannose substituents, is not essential for secretion and expression of enzymatic activity of the transfected gene product. Chapman and Hall, 1992 yeast acid phosphatase nationallicence glycosylation nationallicence transfection nationallicence BHK cells nationallicence APase : acid phosphatase nationallicence PBS : phosphate buffered saline nationallicence TBS : Tris buffered saline nationallicence con A : concanavalin A nationallicence TCA : Tetracarpidium conophorum agglutinin nationallicence Reljic Rajko Laboratory of Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Yugoslavia aut Barbaric Slobodan Laboratory of Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Yugoslavia aut Ries Blanka Laboratory of Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Yugoslavia aut Buxton Roger National Institute for Medical Research, The Ridgeway, Mill Hill, NW71AA, London, UK aut Hughes R. National Institute for Medical Research, The Ridgeway, Mill Hill, NW71AA, London, UK aut Glycoconjugate Journal Kluwer Academic Publishers 9/1(1992-02-01), 39-44 0282-0080 9:1<39 1992 9 10719 https://doi.org/10.1007/BF00731176 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00731176 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Reljic Rajko Laboratory of Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Yugoslavia aut NATIONALLICENCE 700 1- Barbaric Slobodan Laboratory of Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Yugoslavia aut NATIONALLICENCE 700 1- Ries Blanka Laboratory of Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Yugoslavia aut NATIONALLICENCE 700 1- Buxton Roger National Institute for Medical Research, The Ridgeway, Mill Hill, NW71AA, London, UK aut NATIONALLICENCE 700 1- Hughes R. National Institute for Medical Research, The Ridgeway, Mill Hill, NW71AA, London, UK aut NATIONALLICENCE 773 0- Glycoconjugate Journal Kluwer Academic Publishers 9/1(1992-02-01), 39-44 0282-0080 9:1<39 1992 9 10719 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer