Carbohydrate structures of a human tissue plasminogen activator variant expressed in recombinant Chinese hamster ovary cells

FEBS Letters

Noll, Gabriele²  Pâques, Eric-Paul²  Nimtz, Manfred¹  Conradt, Harald S.¹ 

[1] Department of Cell Biology and Genetics, GBF-Gesellschaft für Biotechnologische Forschung mbH, Mascheroder Weg 1, D-3300 Braunschweig, FRG;Research Department, Behringwerke AG, D-3350 Marburg, FRG

关键词: Recombinant glycoprotein;    Structure of N-linked carbohydrates;    Fast atom bombardment mass spectrometry;    Human tissue plasminogen activator;   

DOI  :  10.1016/0014-5793(90)80361-L

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The carbohydrate structures of a genetically engineered human tissue plasminogen activator variant bearing a single N-glycosylation site at Asn 448 are reported. After isolation of the tryptic glycopeptide and liberation of the N-linked carbohydrates by polypeptide:N-glycosidase F, 6 major oligosaccharide fractions were separated by HPLC on NH₂-bonded phase. Their structures were determined by compositional and methylation analyses combined with fast atom bombardment mass spectrometry. Seventy percent of the carbohydrates were of the biantennary complex type with fucose at the proximal GlcNAc and zero, one or two α2-3 linked NeuAc. The remainder were triantennary structures with one, two or three NeuAc.

【 授权许可】

Unknown