PMID: 7016170Apr 14, 1981Paper

Catalytic mechanism of valyl-tRNA synthetase from baker's yeast. Reaction pathway and rate-determining step in the aminoacylation of tRNAVal

Biochemistry
D Kern, J Gangloff

Abstract

The catalytic mechanism of valyl-tRNA synthetase from baker's yeast has been investigated by pre-steady-state and steady-state kinetic measurements and end product dissociation studies. The pre-steady-state kinetics show a lag period during the early time when the reaction is started with free enzyme. The preincubation of the synthetase with tRNAVal and/or valine or preformation of Val approximately AMP leads to a progressive suppression of the lag. This lag probably reflects conformational transitions of the enzyme-substrate complex necessary for the transfer. At low pH or at a low ionic strength, the tRNAVal charging occurs much faster at the pre steady state than at the steady state. We show that after the fast transfer of valine from adenylate to tRNAVal, followed by the fast dissociation of AMP and PPi, a new adenylate is synthesized which promotes the dissociation of the nascent Val-tRNAVal. This dissociation occurs in a multistep process. First ATP and magnesium promote the ejection of the valine moiety of Val-tRNAVal from the adenylate site. A new adenylate is then synthesized which promotes, in the presence of magnesium, several state changes of the end product complex. A complex is finally generated in which the enzym...Continue Reading

References

Dec 1, 1977·Nucleic Acids Research·M SilberklangU L RajBhandary
May 1, 1975·European Journal of Biochemistry·A C CarrJ A Plumbridge
Jan 1, 1979·Annual Review of Biochemistry·P R Schimmel, D Söll
Nov 1, 1976·European Journal of Biochemistry·A DietrichJ P Ebel
Apr 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·F Von Der Haar, E Gaertner
Nov 1, 1974·European Journal of Biochemistry·J RicardJ Buc
Nov 21, 1972·European Journal of Biochemistry·D KernJ P Ebel
Nov 1, 1974·European Journal of Biochemistry·J C MeunierJ Ricard

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Citations

Dec 28, 1981·Biochimica Et Biophysica Acta·E E CreppyG Dirheimer
Mar 29, 1990·Biochimica Et Biophysica Acta·N Menhart, T Viswanatha
Jun 5, 2008·FEMS Microbiology Reviews·Corinne D Hausmann, Michael Ibba
Aug 1, 1982·International Journal of Peptide and Protein Research·F PanH H Lee

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