title: Substrate-Assisted and Enzymatic Pretransfer Editing of Nonstandard Amino Acids by Methionyl-tRNA Synthetase.
abstract: Aminoacyl-tRNA synthetases (aaRSs) are central 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 investigated mechanisms by which methionyl-tRNA synthetase (MetRS) may edit against the highly toxic, noncognate, amino acids homocysteine (Hcy) and its oxygen analogue, homoserine (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.
authors: Department of Chemistry and Biochemistry, University of Windsor , Windsor, Ontario N9B 3P4, Canada.
authors: Grant B
authors: GB
authors: Fortowsky
authors: Department of Chemistry and Biochemistry, University of Windsor , Windsor, Ontario N9B 3P4, Canada.
authors: Daniel J
authors: DJ
authors: Simard
authors: Department of Chemistry and Biochemistry, University of Windsor , Windsor, Ontario N9B 3P4, Canada.
authors: Mohamed M
authors: MM
authors: Aboelnga
authors: Department of Chemistry and Biochemistry, University of Windsor , Windsor, Ontario N9B 3P4, Canada.
authors: James W
authors: JW
authors: Gauld
mesh_terms: D006710:Homocysteine / Q000737:chemistry* / Q000235:genetics; D006714:Homoserine / Q000737:chemistry*; D008718:Methionine-tRNA Ligase / Q000737:chemistry* / Q000235:genetics; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D009154:Mutation; D011485:Protein Binding; D011789:Quantum Theory; D017393:RNA Editing; D013379:Substrate Specificity; D013816:Thermodynamics
chemical_list: D006710:Homocysteine; D006714:Homoserine; D008718:Methionine-tRNA Ligase