Some Surprising Implications of NMR-directed Simulations of Substrate Recognition and Binding by Cytochrome P450_cam (CYP101A1).

ABSTRACT: Cytochrome P450_cam (CYP101A1) catalyzes the stereospecific 5-exo hydroxylation of d-camphor by molecular oxygen. Previously, residual dipolar couplings measured for backbone amide ¹H-¹⁵N correlations in both substrate-free and bound forms of CYP101A1 were used as restraints in soft annealing molecular dynamic simulations in order to identify average conformations of the enzyme with and without substrate bound. Multiple substrate-dependent conformational changes remote from the enzyme active site were identified, and site-directed mutagenesis and activity assays confirmed the importance of these changes in substrate recognition. The current work makes use of perturbation response scanning (PRS) and umbrella sampling molecular dynamic of the residual dipolar

SUBMITTER: Asciutto EK 

PROVIDER: S-EPMC5916322 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature