title: Coupling of GTP hydrolysis by elongation factor G to translocation and factor recycling on the ribosome.
abstract: The translocation step of elongation entails the coordinated movement of tRNA and mRNA on the ribosome. Translocation is promoted by elongation factor G (EF-G) and accompanied by GTP hydrolysis, which affects both translocation and turnover of EF-G. Both reactions are much slower (50-100-fold) when GTP is replaced with non-hydrolyzable GTP analogues or GDP, indicating that the reaction rates are determined by conformational transitions induced by GTP hydrolysis. Compared to the rate of uncatalyzed, spontaneous translocation, ribosome binding of EF-G with any guanine nucleotide reduces the free energy of activation by about 18 kJ/mol, whereas GTP hydrolysis contributes another 10 kJ/mol. The acceleration by GTP hydrolysis is due to large decrease in activation enthalpy by about 30 kJ/mol, compared to the reaction with GTP analogues or GDP, whereas the activation entropy becomes unfavorable and is lowered by about 20 kJ/mol (37 degrees C). The data suggest that GTP hydrolysis induces, by a conformational change of EF-G, a rapid conformational rearrangement of the ribosome ("unlocking") which determines the rates of both tRNA-mRNA translocation and recycling of the factor.
authors.affiliation: Institutes of Molecular Biology and Physical Biochemistry, University of Witten/Herdecke, 58448 Witten, Germany.
authors.forename: Vladimir I
authors.initials: VI
authors.lastname: Katunin
authors.forename: Andreas
authors.initials: A
authors.lastname: Savelsbergh
authors.forename: Marina V
authors.initials: MV
authors.lastname: Rodnina
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wintermeyer
mesh_terms: D002384:Catalysis; D004734:Energy Metabolism; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D020653:Peptide Elongation Factor G / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D012270:Ribosomes / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013873:Thionucleotides / Q000378:metabolism
chemical_list: D020653:Peptide Elongation Factor G; D013873:Thionucleotides; C061507:P(3)-1-(2-nitro)phenylethylguanosine 5'-O-triphosphate; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate