Arginyl-tRNA synthetase from Baker's yeast. Order of substrate addition and action of ATP analogs in the aminoacylation reaction; influence of pyrophosphate on the catalytic mechanism.
The order of substrate addition to arginyl-tRNA synthetase from baker's yeast has been investigated by bisubstrate kinetics, product inhibition and inhibition by three different inhibiting ATP analogs, the 6-N-benzyl, 8-bromo and 3'-deoxy derivatives of ATP, each acting competitively with respect to one of the substrates. The kinetic patterns are consistent with a random ter-ter mechanism, an addition of the three substrates and release of the products in random order. The different inhibitors are bound to different enzyme . substrate complexes of the reaction sequence. Addition of inorganic pyrophosphatase changes the inhibition patterns and addition of methylenediphosphonate as pyrophosphate analog abolishes the effect of pyrophosphatase, showing that the concentration of pyrophosphate is determinant for the mechanism of catalysis.
D000215:Acylation; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives; D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D001126:Arginine-tRNA Ligase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002384:Catalysis; D011756:Diphosphates / Q000494:pharmacology; D007700:Kinetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013379:Substrate Specificity
D011756:Diphosphates; D000255:Adenosine Triphosphate; D000604:Amino Acyl-tRNA Synthetases; D001126:Arginine-tRNA Ligase