Abstract
Enzymes play important roles in many biological processes. Amino acid residues in the active site pocket of an enzyme, which are in direct contact with the substrate(s), are generally believed to be critical for substrate recognition and catalysis. Identifying and understanding how these “catalytic” residues help enzymes achieve enormous rate enhancement has been the focus of many structural and biochemical studies over the past several decades. Recent studies have shown that enzymes are intrinsically dynamic and dynamic coupling between distant structural elements is essential for effective catalysis in modular enzymes. Therefore, distal residues are expected to have impact on enzyme function. However, few studies have investigated the role of distal residues on enzymatic catalysis. In the present study, the effects of distal residue mutations on the catalytic function of an aminoacyl-tRNA synthetase, namely, prolyl-tRNA synthase, were investigated. The present study demonstrates that distal residues significantly contribute to catalysis of the modular Escherichia coli prolyl-tRNA synthetase by maintaining intrinsic protein flexibility.
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The authors confirm that the data supporting the findings of this study within the article [and/or] its supplementary materials will be made available upon request.
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An online software was used for the present study.
Abbreviations
- ARS:
-
Aminoacyl-tRNA synthetase
- Ec:
-
Escherichia coli
- INS:
-
Insertion domain
- PBL:
-
Proline-binding loop
- Pro-AMP:
-
Prolyl-adenylate
- ProRS:
-
Prolyl-tRNA synthetase
- WT:
-
Wild-type
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Funding
This work was supported in part by National Institute of Health [Grant Number: GM117510-01 (S.H.)] and by the Office of Research and Sponsored Programs of the University of Wisconsin-Eau Claire, Eau Claire, WI.
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Zajac, J., Anderson, H., Adams, L. et al. Effects of Distal Mutations on Prolyl-Adenylate Formation of Escherichia coli Prolyl-tRNA Synthetase. Protein J 39, 542–553 (2020). https://doi.org/10.1007/s10930-020-09910-3
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DOI: https://doi.org/10.1007/s10930-020-09910-3