Mechanism of the activation step of the aminoacylation reaction: a significant difference between class I and class II synthetases.
In the present work we report, for the first time, a novel difference in the molecular mechanism of the activation step of aminoacylation reaction between the class I and class II aminoacyl tRNA synthetases (aaRSs). The observed difference is in the mode of nucleophilic attack by the oxygen atom of the carboxylic group of the substrate amino acid (AA) to the αP atom of adenosine triphosphate (ATP). The syn oxygen atom of the carboxylic group attacks the α-phosphorous atom (αP) of ATP in all class I aaRSs (except TrpRS) investigated, while the anti oxygen atom attacks in the case of class II aaRSs. The class I aaRSs investigated are GluRS, GlnRS, TyrRS, TrpRS, LeuRS, ValRS, IleRS, CysRS, and MetRS and class II aaRSs investigated are HisRS, LysRS, ProRS, AspRS, AsnRS, AlaRS, GlyRS, PheRS, and ThrRS. The variation of the electron density at bond critical points as a function of the conformation of the attacking oxygen atom measured by the dihedral angle ψ (C(α)-C') conclusively proves this. The result shows that the strength of the interaction of syn oxygen and αP is stronger than the interaction with the anti oxygen for class I aaRSs. This indicates that the syn oxygen is the most probable candidate for the nucleophilic attack in class I aaRSs. The result is further supported by the computation of the variation of the nonbonded interaction energies between αP atom and anti oxygen as well as syn oxygen in class I and II aaRSs, respectively. The difference in mechanism is explained based on the analysis of the electrostatic potential of the AA and ATP which shows that the relative arrangement of the ATP with respect to the AA is opposite in class I and class II aaRSs, which is correlated with the organization of the active site in respective aaRSs. A comparative study of the reaction mechanisms of the activation step in a class I aaRS (Glutaminyl tRNA synthetase) and in a class II aaRS (Histidyl tRNA synthetase) is carried out by the transition state analysis. The atoms in molecule analysis of the interaction between active site residues or ions and substrates are carried out in the reactant state and the transition state. The result shows that the observed novel difference in the mechanism is correlated with the organizations of the active sites of the respective aaRSs. The result has implication in understanding the experimentally observed different modes of tRNA binding in the two classes of aaRSs.
Department of Chemistry, University of Kalyani, Kalyani, Nadia, West Bengal, 741235, India.
Sindrila Dutta
SD
Banik
Nilashis
N
Nandi
D000255:Adenosine Triphosphate / Q000737:chemistry; D000465:Algorithms; D020816:Amino Acid Motifs; D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry*; D020134:Catalytic Domain; D003198:Computer Simulation; D006643:Histidine-tRNA Ligase / Q000737:chemistry*; D008958:Models, Molecular; D011485:Protein Binding; D012351:RNA, Transfer, Gln / Q000737:chemistry; D012354:RNA, Transfer, His / Q000737:chemistry; D013499:Surface Properties; D013816:Thermodynamics
D012351:RNA, Transfer, Gln; D012354:RNA, Transfer, His; D000255:Adenosine Triphosphate; D000604:Amino Acyl-tRNA Synthetases; C020659:glutaminyl-tRNA synthetase; D006643:Histidine-tRNA Ligase