A comparative study of essential arginine residues in Gramicidin S synthetase 2 and isoleucyl tRNA synthetase.
In gramicidin S synthetase 2 (GS 2) from Bacillus brevis, L-proline, L-valine, L-ornithine, and L-leucine activations to aminoacyl adenylates are progressively inhibited by phenylglyoxal. The inactivation of GS 2 obeys pseudo-first-order kinetics. ATP completely prevents inactivation of GS 2 by phenylglyoxal, whereas amino acids only partially prevent it. In the presence of ATP, four arginine residues per mol of GS 2 are protected from modification by phenylglyoxal as determined by amino acid analysis and the incorporation of [7-14C]phenylgloxal into the enzyme protein, indicating that a single arginine residue is necessary for each amino acid activation. In isoleucyl tRNA synthetase from Escherichia coli, phenylglyoxal inhibits activation of L-isoleucine to isoleucyl adenylate. ATP completely prevents inactivation, although isoleucine only partially prevents it. One arginine residue of isoleucyl tRNA synthetase is protected by ATP from modification by phenylglyoxal, suggesting that a single arginine residue is essential for isoleucine activation. These results support the involvement of arginine residues in ATP binding with GS 2 or isoleucyl tRNA synthetase, and thus indicate that arginine residues of amino acid activating enzymes are essential for the formation of aminoacyl adenylates in both nonribosomal and ribosomal peptide biosynthesis.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000591:Amino Acid Isomerases; D000604:Amino Acyl-tRNA Synthetases; D001120:Arginine / Q000032:analysis*; D001407:Bacillus / Q000201:enzymology; D055598:Chemical Phenomena; D002621:Chemistry; D004926:Escherichia coli / Q000201:enzymology; D007533:Isoleucine-tRNA Ligase / Q000037:antagonists & inhibitors / Q000302:isolation & purification; D009097:Multienzyme Complexes; D010453:Peptide Synthases; D010658:Phenylglyoxal / Q000494:pharmacology; D013379:Substrate Specificity
D009097:Multienzyme Complexes; D000255:Adenosine Triphosphate; D001120:Arginine; D000591:Amino Acid Isomerases; D000604:Amino Acyl-tRNA Synthetases; D007533:Isoleucine-tRNA Ligase; D010453:Peptide Synthases; C020831:gramicidin-S-synthetase 2; D010658:Phenylglyoxal