caa a22 4500 475744098 CHVBK 20180406123508.0 cr unu---uuuuu 170329e20001001xx s 000 0 eng 10.1023/A:1007194101107 doi (NATIONALLICENCE)springer-10.1023/A:1007194101107 Enzyme-Induced Covalent Modification of Methionyl-tRNA Synthetase from Bacillus stearothermophilus by Methionyl-Adenylate: Identification of the Labeled Amino Acid Residues by Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry [Elektronische Daten] [Codjo Hountondji, Christian Beauvallet, Jean-Claude Pernollet, Sylvain Blanquet] Methionyl-tRNA synthetase (MetRS) from Bacillus stearothermophilus was shown to undergo covalent methionylation by a donor methionyl-adenylate, the mixed carboxylic-phosphoric acid anhydride synthesized by the enzyme itself. Covalent reaction of methionyl-adenylate with the synthetase or other proteins proceeds through the formation of an isopeptide bond between the carboxylate of the amino acid and the ∈-NH2 group of lysyl residues. The stoichiometries of labeling, as followed by TCA precipitation, were 2.2 ± 0.1 and 4.3 ± 0.1 mol of [14C]Met incorporated by 1 mol of the monomeric MS534 and the native dimeric species of B. stearo methionyl-tRNA synthetase, respectively. Matrix-assisted laser desorption-ionization mass spectrometry designated lysines-261, -295, -301 and -528 (or -534) of truncated methionyl-tRNA synthetase as the target residues for covalent binding of methionine. By analogy with the 3D structure of the monomeric M547 species of E. coli methionyl-tRNA synthetase, lysines-261, -295, and -301 would be located in the catalytic crevice of the thermostable enzyme where methionine activation and transfer take place. It is proposed that, once activated by ATP, most of the methionine molecules react with the closest reactive lysyl residues. Plenum Publishing Corporation, 2000 Methionyl-tRNA synthetase nationallicence methionyl-adenylate nationallicence isopeptide bond nationallicence MALDI-MS nationallicence posttranslational modification nationallicence Hountondji Codjo Ecole Polytechnique, Laboratoire de Biochimie (CNRS UMR 7654), 91128, Palaiseau Cédex, France aut Beauvallet Christian Unité de Recherches en Biochimie et Structure des Protéines, Domaine de Vilvert, Institut National de la Recherche Agronomique (INRA), 78352, Jouy-en-Josas Cedex, France aut Pernollet Jean-Claude Unité de Recherches en Biochimie et Structure des Protéines, Domaine de Vilvert, Institut National de la Recherche Agronomique (INRA), 78352, Jouy-en-Josas Cedex, France aut Blanquet Sylvain Ecole Polytechnique, Laboratoire de Biochimie (CNRS UMR 7654), 91128, Palaiseau Cédex, France aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/7(2000-10-01), 563-568 0277-8033 19:7<563 2000 19 10930 https://doi.org/10.1023/A:1007194101107 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1007194101107 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hountondji Codjo Ecole Polytechnique, Laboratoire de Biochimie (CNRS UMR 7654), 91128, Palaiseau Cédex, France aut NATIONALLICENCE 700 1- Beauvallet Christian Unité de Recherches en Biochimie et Structure des Protéines, Domaine de Vilvert, Institut National de la Recherche Agronomique (INRA), 78352, Jouy-en-Josas Cedex, France aut NATIONALLICENCE 700 1- Pernollet Jean-Claude Unité de Recherches en Biochimie et Structure des Protéines, Domaine de Vilvert, Institut National de la Recherche Agronomique (INRA), 78352, Jouy-en-Josas Cedex, France aut NATIONALLICENCE 700 1- Blanquet Sylvain Ecole Polytechnique, Laboratoire de Biochimie (CNRS UMR 7654), 91128, Palaiseau Cédex, France aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/7(2000-10-01), 563-568 0277-8033 19:7<563 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer