Carbohydrate structures of a human tissue plasminogen activator variant expressed in recombinant Chinese hamster ovary cells.
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 alpha 2-3 linked NeuAc. The remainder were triantennary structures with one, two or three NeuAc.
Nimtz
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Department of Cell Biology and Genetics, GBF-Gesellschaft für Biotechnologische Forschung mbH, Braunschweig, FRG.
Noll
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G
Pâques
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EP
Conradt
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HS
D000818:Animals; D001216:Asparagine / Q000235:genetics; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002460:Cell Line; D002999:Clone Cells; D006224:Cricetinae; D003412:Cricetulus; D014644:Genetic Variation; D005973:Glutamine / Q000235:genetics; D006023:Glycoproteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D006031:Glycosylation; D006801:Humans; D008745:Methylation; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*; D010960:Plasminogen Activators / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins
D006023:Glycoproteins; D009844:Oligosaccharides; D011994:Recombinant Proteins; D005973:Glutamine; D001216:Asparagine; D010960:Plasminogen Activators