Document Type

Article

Publication Date

2015

Publication Title

Biochemistry

Volume

54

First Page

5757

Last Page

5765

DOI

10.1021/acs.biochem.5b00588

Keywords

Computational chemistry, enzymology, aminoacyl-tRNA Synthetases, editing, in silico, mutagensis

Abstract

Aminoacyl-tRNA synthetases (aaRSs) are cen- tral to a number of physiological processes, including protein biosynthesis. In particular, they activate and then transfer their corresponding amino acid to the cognate tRNA. This is achieved with a generally remarkably high fidelity by editing against incorrect standard and nonstandard amino acids. Using docking, molecular dynamics (MD), and hybrid quantum mechanical/molecular mechanics methods, we have inves- tigated mechanisms by which methionyl-tRNA synthetase (MetRS) may edit against the highly toxic, noncognate, amino acids homocysteine (Hcy) and its oxygen analogue, homo- serine (Hse). Substrate-assisted editing of Hcy-AMP in which its own phosphate acts as the mechanistic base occurs with a rate-limiting barrier of 98.2 kJ mol−1. This step corresponds to nucleophilic attack of the Hcy side-chain sulfur at its own carbonyl carbon (CCarb). In contrast, a new possible editing mechanism is identified in which an active site aspartate (Asp259) acts as the base. The rate-limiting step is now rotation about the substrate’s aminoacyl Cβ−Cγ bond with a barrier of 27.5 kJ mol−1, while for Hse-AMP, the rate-limiting step is cleavage of the CCarb−OP bond with a barrier of 30.9 kJ mol−1. A similarly positioned aspartate or glutamate also occurs in the homologous enzymes LeuRS, IleRS, and ValRS, which also discriminate against Hcy. Docking and MD studies suggest that at least in the case of LeuRS and ValRS, a similar editing mechanism may be possible.

Comments

This article was fist published in Biochemistry and can be downloaded here: http://pubs.acs.org/doi/10.1021/acs.biochem.5b00588

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