title: Magnesium dependence of the measured equilibrium constants of aminoacyl-tRNA synthetases.
abstract: The apparent equilibrium constants (K') for six reactions catalyzed by aminoacyl-tRNA synthetases from Escherichia coli were measured, the equations for the magnesium dependence of the equilibrium constants were derived, and best-fit analyses between the measured and calculated values were used. The K' values at 1 mM Mg(2+) ranged from 0.49 to 1.13. The apparent equilibrium constants increased with increasing Mg(2+) concentrations. The values were 2-3 times higher at 20 mM Mg(2+) than at 1 mM Mg(2+), and the dependence was similar in the class I and class II synthetases. The main reason for the Mg(2+) dependence is the existence of PP(i) as two magnesium complexes, but only one of them is the real product. AMP exists either as free AMP or as MgAMP, and therefore also has some effect on the measured equilibrium constant. However, these dependences alone cannot explain the measured results. The measured dependence of the K' on the Mg(2+) concentration is weaker than that caused by PP(i) and AMP. Different bindings of the Mg(2+) ions to the substrate tRNA and product aminoacyl-tRNA can explain this observation. The best-fit analysis suggests that tRNA reacts as a magnesium complex in the forward aminoacylation direction but this given Mg(2+) ion is not bound to aminoacyl-tRNA at the start of the reverse reaction. Thus Mg(2+) ions seem to have an active catalytic role, not only in the activation of the amino acid, but in the posttransfer steps of the aminoacyl-tRNA synthetase reaction, too.
authors: Department of Biochemistry, University of Turku, FIN-20014, Turku, Finland. kalervo.airas@utu.fi
authors: R Kalervo
authors: RK
authors: Airas
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D002384:Catalysis; D004926:Escherichia coli / Q000201:enzymology*; D008274:Magnesium / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D000604:Amino Acyl-tRNA Synthetases; D008274:Magnesium