Affordable Access

deepdyve-link
Publisher Website

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

Authors
  • Kaesler, Nadine
  • Schettgen, Thomas
  • Mutucumarana, Vasantha P.
  • Brandenburg, Vincent
  • Jahnen-Dechent, Willi
  • Schurgers, Leon J.
  • Krüger, Thilo
Type
Published Article
Journal
Analytical Biochemistry
Publication Date
Jan 01, 2011
Accepted Date
Nov 28, 2011
Volume
421
Issue
2
Pages
411–416
Identifiers
DOI: 10.1016/j.ab.2011.11.036
Source
Elsevier
Keywords
License
Unknown

Abstract

The gamma (γ)-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 γ-carboxyglutamic acid residues in proteins. The activity of tissue derived γ-glutamyl carboxylase is commonly assayed by incorporation of H14CO3- 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 γ-glutamyl carboxylated form by reversed-phase HPLC. This method offers a convenient alternative to the established radioactive labeling techniques.

Report this publication

Statistics

Seen <100 times