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Carbohydrate structures of a human tissue plasminogen activator variant expressed in recombinant Chinese hamster ovary cells
Author(s) -
Nimtz Manfred,
Noll Gabriele,
Pâques Eric-Paul,
Conradt Harald S.
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)80361-l
Subject(s) - chinese hamster ovary cell , fucose , glycopeptide , chemistry , carbohydrate , biochemistry , glycosylation , oligosaccharide , plasminogen activator , recombinant dna , glycoprotein , glycoside hydrolase , glycan , activator (genetics) , fucosylation , mannose , microbiology and biotechnology , enzyme , biology , receptor , gene , antibiotics , endocrinology
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 NH 2 ‐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.