Identification of the vitamin K-dependent carboxylase active site: Cys-99 and Cys-450 are required for both epoxidation and carboxylation -- Data Supplement - Supplemental Data -- Proceedings of the National Academy of Sciences

Supplementary material for Pudota et al. (2000) Proc. Natl. Acad. Sci. USA 97 (24), 13033–13038.

Fig. 4.
Time- and dose-dependent NEM inhibition of carboxylation and epoxidation. (A) Freshly prepared pure carboxylase (20 µl, 0.15 µM), purified in the absence of reducing agents, was incubated with 10 mM NEM for 0–30 min, followed by quenching with excess DTT and assaying for carboxylase or epoxidase activities (150 µl cocktail, 60 min). (B) Aliquots from the same carboxylase preparation (20 µl, 0.15 µM) were incubated with 0–20 mM NEM for 30 min, then quenched with DTT and assayed as in A.