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

FEBS Letters

Vo, Hung C²  Chen, David D.Y¹  Britz-Mckibbin, Philip¹  MacGillivray, Ross T.A² 

[1] Department of Chemistry, University of British Columbia, Vancouver, B.C. V6T 1Z3, Canada;Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, B.C. V6T 1Z3, Canada

关键词: Gamma carboxy glutamic acid;    Prothrombin;    Capillary electrophoresis;    Laser-induced fluorescence;    Recombinant protein;    Baby hamster kidney cell;   

DOI  :  10.1016/S0014-5793(99)00131-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The γ-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.

【 授权许可】

Unknown