On the composition and function of the carbohydrate moiety of tissue-type plasminogen activator from human melanoma cells

Two variants (I and II) of tissue-type plasminogen activator (t-PA) from human melanoma cells were separated by Lysine Sepharose chromatography. The carbohydrate compositions of the forms were determined by gas-liquid chromatography. Variant I contained 12.8 g and variant II 7.1 g of carbohydrate pe...

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Bibliographic Details
Published inThrombosis and haemostasis Vol. 54; no. 4; p. 788
Main Authors Rijken, D C, Emeis, J J, Gerwig, G J
Format Journal Article
LanguageEnglish
Published Germany 1985
Subjects
Carbohydrates - analysis
Cells, Cultured
Chromatography, Agarose
Fibrin - metabolism
Humans
Melanoma - metabolism
Tissue Plasminogen Activator - isolation & purification
Tissue Plasminogen Activator - metabolism
Tunicamycin - pharmacology
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Summary:Two variants (I and II) of tissue-type plasminogen activator (t-PA) from human melanoma cells were separated by Lysine Sepharose chromatography. The carbohydrate compositions of the forms were determined by gas-liquid chromatography. Variant I contained 12.8 g and variant II 7.1 g of carbohydrate per 100 g protein. Both variants contained N-acetylgalactosamine, suggesting O-glycosylation in addition to N-glycosylation. The possible role of N-linked oligosaccharides for the biological activity of t-PA was studied using t-PA secreted by melanoma cells in the presence of tunicamycin, an inhibitor of N-glycosylation. The latter t-PA showed the same plasminogen activating and fibrin binding properties as normally glycosylated t-PA, indicating that N-linked carbohydrate is not involved in the fibrinolytic activity of t-PA.
ISSN:0340-6245
DOI:10.1055/s-0038-1660134