title: Effector region of the translation elongation factor EF-Tu.GTP complex stabilizes an orthoester acid intermediate structure of aminoacyl-tRNA in a ternary complex.
abstract: tRNA(Val) from Escherichia coli was aminoacylated with [1-13C]valine and its complex with Thermus thermophilus elongation factor EF-Tu.GTP was analyzed by 13C NMR spectroscopy. The results suggest that the aminoacyl residue of the valyl-tRNA in ternary complex with bacterial EF-Tu and GTP is not attached to tRNA by a regular ester bond to either a 2'- or 3'-hydroxyl group; instead, an intermediate orthoester acid structure with covalent linkage to both vicinal hydroxyls of the terminal adenosine-76 is formed. Mutation of arginine-59 located in the effector region of EF-Tu, a conserved residue in protein elongation factors and the alpha subunits of heterotrimeric guanine nucleotide-binding regulatory proteins (G proteins), abolishes the stabilization of the orthoester acid structure of aminoacyl-tRNA.
authors.affiliation: Laboratorium für Biochemie, Universität Bayreuth, Federal Republic of Germany.
authors.forename: C
authors.initials: C
authors.lastname: Förster
authors.forename: S
authors.initials: S
authors.lastname: Limmer
authors.forename: W
authors.initials: W
authors.lastname: Zeidler
authors.forename: M
authors.initials: M
authors.lastname: Sprinzl
mesh_terms: D001665:Binding Sites; D002247:Carbon Isotopes; D004926:Escherichia coli / Q000201:enzymology; D004952:Esters; D006160:Guanosine Triphosphate / Q000737:chemistry* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D010444:Peptide Elongation Factor Tu / Q000737:chemistry* / Q000378:metabolism; D012366:RNA, Transfer, Val / Q000737:chemistry* / Q000378:metabolism; D016963:Thermus thermophilus / Q000378:metabolism*; D014637:Valine-tRNA Ligase / Q000378:metabolism
chemical_list: D002247:Carbon Isotopes; D004952:Esters; D012366:RNA, Transfer, Val; D006160:Guanosine Triphosphate; D010444:Peptide Elongation Factor Tu; D014637:Valine-tRNA Ligase