Peptide mapping of aminoacyl-tRNA synthetases: evidence for internal sequence homology in Escherichia coli leucyl-tRNA synthetase.
Most aminoacyl-tRNA synthetases contain polypeptide chains of about either 50,000 or 100,000 daltons. Peptide mapping of tryptic, chymotryptic, or Staphylococcus aureus acid protease digests of seryl-tRNA synthetase (100,000, dimer) and leucyl-tRNA synthetase (100,000, monomer) from E. coli was done after selective modification of lysine residues with [(14)C]succinic anhydride or of methionine residues with [(14)C]iodoacetate. By use of thin-layer electrophoresis and chromatography on silicagel or cellulose plates followed by radioautography it was possible, depending upon the specific activity of the reagent used, to detect radioactive peptides obtained from as little as l mug of protein.Seryl-tRNA synthetase gave the correct number of tryptic peptides expected for a dimer of identical subunits. Leucyl-tRNA synthetase, on the other hand, gave roughly half the number of radioactive tryptic, chymotryptic, and acid protease peptides expected from the lysine, arginine, and methionine content of the 100,000 monomer. We have interpreted these results as indicating that extensive internal homology exists among lysine- and methionine-containing peptides within the leucyl-tRNA synthetase. The simplest conclusion that can be drawn from these observations is that the NH(2)- and COOH-terminal halves of leucyl-tRNA synthetase and perhaps other synthetases of 100,000 molecular weight may have evolved through a process of gene duplication and fusion, followed by limited diversification by way of amino-acid substitutions accumulating during evolution.
D000604:Amino Acyl-tRNA Synthetases / Q000032:analysis*; D000812:Anhydrides; D001345:Autoradiography; D002250:Carbon Radioisotopes; D002845:Chromatography; D002918:Chymotrypsin; D004586:Electrophoresis; D004926:Escherichia coli / Q000201:enzymology*; D006868:Hydrolysis; D007461:Iodoacetates; D007930:Leucine; D008970:Molecular Weight; D010447:Peptide Hydrolases; D010455:Peptides / Q000032:analysis; D013210:Staphylococcus / Q000201:enzymology; D013386:Succinates; D014357:Trypsin
D000812:Anhydrides; D002250:Carbon Radioisotopes; D007461:Iodoacetates; D010455:Peptides; D013386:Succinates; D010447:Peptide Hydrolases; D002918:Chymotrypsin; D014357:Trypsin; D000604:Amino Acyl-tRNA Synthetases; D007930:Leucine