Global Kinetic Mechanism of Microsomal Glutathione Transferase 1 and Insights into Dynamic Enzyme Activation.

ABSTRACT: Microsomal glutathione transferase 1 (MGST1) has a unique ability to be activated, ≤30-fold, by modification with sulfhydryl reagents. MGST1 exhibits one-third-of-the-sites reactivity toward glutathione and hence heterogeneous binding to different active sites in the homotrimer. Limited turnover stopped-flow kinetic measurements of the activated enzyme allowed us to more accurately determine the K_D for the "third" low-affinity GSH binding site (1.4 ± 0.3 mM). The rate of thiolate formation, k₂ (0.77 ± 0.06 s^-1), relevant to turnover, could also be determined. By deriving the steady-state rate equation for a random sequential mechanism for MGST1, we can predict K_M, k_cat, and k_cat/K_M values from these and previously

SUBMITTER: Spahiu L 

PROVIDER: S-EPMC5954419 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature