The stereochemical course of the ribosome-dependent GTPase reaction of elongation factor G from Escherichia coli

J Biol Chem. 1981 Aug 10;256(15):7734-7.

Abstract

The stereochemical course of the ribosome-dependent GTPase reaction of elongation factor G from Escherichia coli has been determined. Guanosine 5'-(gamma-thio)triphosphate stereospecifically labeled with 17O and 18O in the gamma-position was hydrolyzed in the presence of the elongation factor and ribosomes. The configuration of the product, inorganic [16O, 17O, 18O]thiophosphate ws analyzed by 31P NMR after its stereospecific incorporation into adenosine 5'-(beta-thio)triphosphate. The analysis showed that the hydrolysis proceeds with inversion of configuration at the transferred phosphorus atom. It is therefore likely that the hydrolysis occurs in a single step by direct, in-line transfer of the phosphorus from GDP to a water oxygen, without a phosphoenzyme intermediate.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphate / analogs & derivatives
  • Adenosine Triphosphate / analysis
  • Escherichia coli / enzymology*
  • GTP Phosphohydrolase-Linked Elongation Factors / metabolism*
  • Magnetic Resonance Spectroscopy
  • Peptide Elongation Factor G
  • Peptide Elongation Factors / metabolism*
  • Phosphoric Monoester Hydrolases / metabolism*
  • Ribosomes / enzymology*
  • Stereoisomerism
  • Substrate Specificity

Substances

  • Peptide Elongation Factor G
  • Peptide Elongation Factors
  • adenosine 5'-O-(3-thiotriphosphate)
  • Adenosine Triphosphate
  • Phosphoric Monoester Hydrolases
  • GTP Phosphohydrolase-Linked Elongation Factors