Crowder-induced Conformational Ensemble Shift in Escherichia Coli Prolyl-tRNA Synthetase

摘要: ABSTRACT The effect of macromolecular crowding on the structure and function Escherichia coli prolyl-tRNA synthetase (Ec ProRS) has been investigated using a combined experimental theoretical method. Ec ProRS is multi-domain enzyme; coupled-domain dynamics essential for efficient catalysis. To gain an insight into mechanistic detail effect, kinetic studies were conducted with varying concentrations sizes crowders. In parallel, spectroscopic quantum chemical employed to probe “soft-interactions” between crowders protein side chains. Finally, dimeric was examined in presence long-duration (70 ns) classical molecular dynamic simulations. results simulations revealed significant shift conformational ensemble, which consistent model explained observed alteration parameters. Collectively, present study demonstrated that effects both catalytic function, are correlated. This first report where found impact ensemble ProRS, member aminoacyl-tRNA family, central synthesis all living cells. affirmed should be considered while investigating structure-dynamics-function relationship modular enzymes.