Comparison of two reducing agents dithiothreitol and tris(3-hydroxypropyl)phosphine for in vitro kinetic assay of vitamin K epoxide reductase.

ABSTRACT: Vitamin K epoxide reductase (VKOR) is a target enzyme for anticoagulants, such as warfarin, that are used as medicines or rodenticides. Assessing VKOR activity is required to ensure the proper usage of these drugs. Dithiothreitol (DTT) is a typical disulfide reductant that is used as a substrate for in vitro VKOR assays. However, DTT is considered problematic because of its side effects. Tris(3-hydroxypropyl)phosphine (THP) has been found to be a reliable alternative to DTT, as shown by kinetic analyses of the VKOR with them. THP showed significantly lower V _max and K_m values than those of DTT; however, there was no significant difference in their V _max/K_m and IC₅₀ for warfarin.

SUBMITTER: Takeda K 

PROVIDER: S-EPMC7386777 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature