Effector region of the translation elongation factor EF-Tu.GTP complex stabilizes an orthoester acid intermediate structure of aminoacyl-tRNA in a ternary complex.
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.
Förster
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C
Laboratorium für Biochemie, Universität Bayreuth, Federal Republic of Germany.
Limmer
S
S
Zeidler
W
W
Sprinzl
M
M
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
D002247:Carbon Isotopes; D004952:Esters; D012366:RNA, Transfer, Val; D006160:Guanosine Triphosphate; D010444:Peptide Elongation Factor Tu; D014637:Valine-tRNA Ligase