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Undercarboxylation of recombinant prothrombin revealed by analysis of gamma-carboxyglutamic acid using capillary electrophoresis and laser-induced fluorescence

TitleUndercarboxylation of recombinant prothrombin revealed by analysis of gamma-carboxyglutamic acid using capillary electrophoresis and laser-induced fluorescence
Publication TypeJournal Article
Year of Publication1999
AuthorsVo, HC, Britz-McKibbin, P, Chen, DDY, MacGillivray, RTA
JournalFebs Letters
Volume445
Pagination256-260
Date PublishedFeb
Type of ArticleArticle
ISBN Number0014-5793
Keywordsbaby hamster kidney, calcium, capillary electrophoresis, cell, EXPRESSION, FACTOR-X, gamma carboxy glutamic acid, GLUTAMIC-ACID, HUMAN PROTEIN-C, LASER-INDUCED FLUORESCENCE, METAL, PHOSPHOLIPID-BINDING-PROPERTIES, PROTHROMBIN, PURIFICATION, recombinant protein, RESIDUES, VITAMIN-K
Abstract

The gamma-carboxyglutamic acid (Gla) content of several variants of human prothrombin has been measured by using capillary electrophoresis and laser-induced fluorescence (CE-LIF). Both plasma-derived prothrombin and recombinant prothrombin contain ten residues of Gla per molecule of protein. In contrast, a variant of human prothrombin (containing the second kringle domain of bovine prothrombin) was separated into two populations that differed in their Gla content. Direct measurement of the Gla content showed an association with the presence or absence of the calcium-dependent conformational change that is required for prothombinase function. Thus, the CE-LIF assay is useful in determining the carboxylation status of recombinant proteins. (C) 1999 Federation of European Biochemical Societies.

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