Seven mammalian aminoacyl-tRNA synthetases associated within the same complex are functionally independent.
A heterotypic multienzyme complex from sheep liver containing seven aminoacyl-tRNA synthetases specific for isoleucine, leucine, methionine, glutamine, glutamic acid, lysine and arginine was subjected to kinetic analyses to examine the possibility that association of these enzymes may impart kinetic properties which differ from those of their unassociated counterparts. The evidence obtained by two different approaches leads to the conclusion that the associated enzymes are functionally independent. Firstly, the kinetic constants of the methionyl-tRNA and lysyl-tRNA synthetase components of the complex do not differ significantly from those of their unassociated counterparts obtained after controlled proteolysis of the complex. Secondly, the methionyl-tRNA synthetase component of the complex displays identical kinetic constants, whether assayed in the presence of [14C]methionine, ATP and highly enriched tRNAMet alone, or in the additional presence of the substrates required for unlabeled aminoacyl-tRNA formation by each of the other six enzymes. Similarly, the initial rates of [14C]aminoacyl-tRNA formation catalyzed by any of the six other enzymes was unaffected by the concomitant functioning of the other aminoacyl-tRNA synthetases. The sedimentation behaviour of the aminoacyl-tRNA synthetase components of the complex under conditions prevailing in the tRNA aminoacylation assay indicates that they remain associated under these conditions. The implications of these findings on the structural organization of the enzymes within the complex are discussed.
D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D000818:Animals; D002499:Centrifugation, Density Gradient; D008099:Liver / Q000201:enzymology; D008250:Lysine-tRNA Ligase / Q000378:metabolism; D008718:Methionine-tRNA Ligase / Q000378:metabolism; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000378:metabolism*; D012756:Sheep; D013379:Substrate Specificity
D009097:Multienzyme Complexes; D000604:Amino Acyl-tRNA Synthetases; D008718:Methionine-tRNA Ligase; D008250:Lysine-tRNA Ligase