Aminoacylation reaction in the histidyl-tRNA synthetase: fidelity mechanism of the activation step.
Aminoacylation is a vital step of natural biosynthesis of peptide. Correct aminoacylation is a necessary prerequisite for the elimination of noncognate amino acids such as D-amino acids. In the present work, we studied the fidelity mechanism of histidine (His) activation (first step of aminoacylation reaction) using a combined quantum mechanical/semiempirical method based on a model of crystal structure of the oligomeric complex of histidyl-tRNA synthetase (HisRS) from Escherichia coli. The study of the variation in the energy during the mutual approach of the His and ATP to form adenylate shows that the surrounding nanospace of synthetase confines the reactants (L-His and ATP) and proximally places in a geometry suitable for the in-line nucleophilic attack. The significantly higher energy of the energy surface of the model containing D-His is due to unfavorable interaction of D-His with ATP and surrounding residues. This indicates that the network of interaction (principally electrostatic) is highly unfavorable when D-amino acid is incorporated. The reorganization of the surrounding nanospace can lower the unfavorable nature of the intermolecular energy surface of D-His and surrounding residues. However, such a rearrangement requires large-scale structural reorganization of the synthetase structure and is unfavorable. The variation in the bond angles and distances in going from the reactant state to the product state via transition state confirms the mechanism of nucleophilic attack and concomitant inversion of oxygen atoms around alpha-phosphorus (alpha-P). Calculation of the electrostatic potential indicates that in addition to the Mg(2+) the Arg residues in the active site facilitate the nucleophilic attack by reducing the negative charge distributed over the oxygen atoms attached to the alpha-P of ATP. Arg 259 residue has a role similar to that played by the two Mg(2+) cations as this residue is in close proximity of the alpha-P of ATP. Arg 113 also facilitates the reduction of the negative charge on the other side of the reaction center. The favorable electrostatic interaction of the Arg 259 with ATP and His is also concluded from the calculation of the binding energy. The Arg 259 anchors the carboxylic acid group of His and the oxygen atom of the alpha-phosphate group during the progress of reaction. Consequently, Arg 259 plays an important catalytic role in the activation step rather than merely reducing the negative charge density over the ATP.
Banik
S Dutta
SD
Department of Chemistry, University of Kalyani, Kalyani, Nadia, West Bengal, 741235, India.
Nandi
N
N
D049148:Aminoacylation; D001665:Binding Sites; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006639:Histidine / Q000737:chemistry; D006643:Histidine-tRNA Ligase / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D055672:Static Electricity
D006639:Histidine; D006643:Histidine-tRNA Ligase