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Undercarboxylation of recombinant prothrombin revealed by analysis of γ‐carboxyglutamic acid using capillary electrophoresis and laser‐induced fluorescence
Author(s) -
Vo Hung C,
Britz-Mckibbin Philip,
Chen David D.Y,
MacGillivray Ross T.A
Publication year - 1999
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/s0014-5793(99)00131-3
Subject(s) - chemistry , recombinant dna , fluorescence , capillary electrophoresis , chromatography , electrophoresis , biochemistry , gene , physics , quantum mechanics
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.