Macromolecular crowding in the cytosolic environment is proposed to influence substrate binding affinity and enzymatic activity of proteins. However, the mechanism of dynamic changes that occur during substrate binding and catalysis in the presence of crowders has remained underexplored for modular enzymes like aminoacyl-tRNA synthetases(AARSs). AARSs catalyze the ligation of an amino acid onto its cognate tRNA, an important step in protein synthesis in all living cells. Currently, we are investigating the impact of macromolecular crowding on the conformational dynamics of prolyl-tRNA synthetases (ProRSs), a member of AARSs family, using intrinsic tryptophan fluorescence measurements and molecular dynamic simulations. We are using synthetic crowding agents to mimic the cytosolic crowding conditions. The impact of both charge and size of crowding agents are being investigated. Herein, we will present the preliminary results of our study, which suggest that crowding agents have significant impact on the conformational dynamics and function of bacterial ProRSs. This knowledge can potentially be used to further our understanding of intracellular enzyme dynamics, as well as to develop selective drugs against pathogenic AARSs.

Exploring the effects of macromolecular crowding on conformational change in <i>escherichia coli</i> prolyl-tRNA synthetase using intrinsic tryptophan fluorescence

2019 Spring National Meeting

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Exploring the effects of macromolecular crowding on conformational change in escherichia coli prolyl-tRNA synthetase using intrinsic tryptophan fluorescence