Thermal stabilization of dihydrofolate reductase using monte carlo unfolding simulations and its functional consequences.

ABSTRACT: Design of proteins with desired thermal properties is important for scientific and biotechnological applications. Here we developed a theoretical approach to predict the effect of mutations on protein stability from non-equilibrium unfolding simulations. We establish a relative measure based on apparent simulated melting temperatures that is independent of simulation length and, under certain assumptions, proportional to equilibrium stability, and we justify this theoretical development with extensive simulations and experimental data. Using our new method based on all-atom Monte-Carlo unfolding simulations, we carried out a saturating mutagenesis of Dihydrofolate Reductase (DHFR), a key target of antibiotics and chemotherapeutic drugs. The method predicted more than 500 stabilizing mutati

SUBMITTER: Tian J 

PROVIDER: S-EPMC4407897 | biostudies-literature | 2015 Apr

REPOSITORIES: biostudies-literature