Interactions of aminoacyl-tRNA synthetases in high-molecular-weight multienzyme complexes from rat liver.
The functional interaction of Arg-, Ile-, Leu-, Lys- and Met-tRNA synthetases occurring within the same rat liver multienzyme complex are investigated by examining the enzymes catalytic activities and inactivation kinetics. The Michaelis constants for amino acids, ATP and tRNAs of the dissociated aminoacyl-tRNA synthetases are not significantly different from those of the high-Mr multienzyme complex, except in a few cases where the Km values of the dissociated enzymes are higher than those of the high-Mr form. The maximal aminoacylation velocities of the individual aminoacyl-tRNA synthetases are not affected by the presence of simultaneous aminoacylation by another synthetase occurring within the same multienzyme complex. Site-specific oxidative modification by ascorbate and nonspecific thermal inactivation of synthetases in the purified rat liver 18 S synthetase complex are examined. Lys- and Arg-tRNA synthetases show remarkably parallel time-courses in both inactivation processes. Leu- and Met-tRNA synthetases also show parallel kinetics in thermal inactivation and possibly oxidative inactivation. Ile-tRNA synthetase shows little inactivation in either process. The oxidative inactivation of Lys- and Arg-tRNA synthetases can be reversed by addition of dithiothreitol. These results suggest that synthetases within the same high-Mr complex catalyze aminoacylation reactions independently; however, the stabilities of some of the synthetases in the multienzyme complex are coupled. In particular, the stability of Arg-tRNA synthetase depends appreciably on its association with fully active Lys-tRNA synthetase.
D000596:Amino Acids / Q000494:pharmacology; D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D000818:Animals; D001126:Arginine-tRNA Ligase / Q000378:metabolism; D001205:Ascorbic Acid / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D006358:Hot Temperature; D007533:Isoleucine-tRNA Ligase / Q000378:metabolism; D007700:Kinetics; D007935:Leucine-tRNA Ligase / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008250:Lysine-tRNA Ligase / Q000378:metabolism; D008718:Methionine-tRNA Ligase / Q000378:metabolism; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000378:metabolism*; D051381:Rats
D000596:Amino Acids; D009097:Multienzyme Complexes; D000604:Amino Acyl-tRNA Synthetases; D008718:Methionine-tRNA Ligase; D001126:Arginine-tRNA Ligase; D007935:Leucine-tRNA Ligase; D007533:Isoleucine-tRNA Ligase; D008250:Lysine-tRNA Ligase; D001205:Ascorbic Acid; D004229:Dithiothreitol