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Effects of Distal Mutations on Prolyl-Adenylate Formation of Escherichia coli Prolyl-tRNA Synthetase

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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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Data Availability

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.

Code Availability

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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Authors and Affiliations

  1. Department of Chemistry, University of Wisconsin-Eau Claire, Eau Claire, USA

    Jonathan Zajac, Heidi Anderson, Lauren Adams, Dechen Wangmo, Shanzay Suhail, Aimee Almen, Lauren Berns, Breanna Coerber, Logan Dawson, Andrea Hunger, Julia Jehn, Joseph Johnson, Naomi Plack, Steven Strasser, Murphi Williams, Sudeep Bhattacharyya & Sanchita Hati

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  1. Jonathan Zajac
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Correspondence to Sanchita Hati.

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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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