A fluorescent method to determine vitamin K-dependent gamma-glutamyl carboxylase activity

作者：Kaesler Nadine; Schettgen Thomas; Mutucumarana Vasantha P; Brandenburg Vincent; Jahnen Dechent Willi; Schurgers Leon J; Krueger Thilo

来源：Analytical Biochemistry, 2012, 421(2): 411-416.

DOI：10.1016/j.ab.2011.11.036

• gamma-Glutamyl carboxylase
• Enzyme activity
• HPLC
• Vitamin K cycle

The gamma (gamma)-glutamyl carboxylase is a key enzyme in vitamin K-dependent carboxylation of proteins involved in hemostasis and inflammation. It is an endoplasmic enzyme posttranslationally converting glutamic acid residues into gamma-carboxyglutamic acid residues in proteins. The activity of tissue derived gamma-glutamyl carboxylase is commonly assayed by incorporation of (HCO3-)-C-14 into synthetic peptides and subsequent quantification using liquid scintillation counting. We present a nonradioactive assay using a fluorescein isothiocyanate-labeled short peptide that can be readily detected in its unmodified and gamma-glutamyl carboxylated form by reversed-phase HPLC. This method offers a convenient alternative to the established radioactive labeling techniques.

• 出版日期2012-2-15