PMID: 7013799Mar 31, 1981Paper

Mechanistic deductions from isotope effects in multireactant enzyme mechanisms

Biochemistry
P F Cook, W W Cleland

Abstract

In the enzymatic mechanism with two or more substrates, comparison of the isotope effects on the maximum velocity and on the apparent V/K values when each substrate concentration is varied allows one to deduce the kinetic mechanism and obtain quantitative information on the relative rates at which substrates dissociate from the enzyme, as opposed to undergoing reaction to give products. Theory is also presented for using the effects of other reactants on the apparent isotope effects determined by the equilibrium perturbation method to determine the same information. With liver alochol dehydrogenase, DPN is not released at an appreciable rate from the E-DPN-cyclohexanol complex, while cyclohexanol is released much more rapidly than it reacts to give products, so that the mechanism appears ordered. With DPNH and cyclohexanone, however, the reaction is random since DPNH can be released from the ternary complex at a finite rate. With yeast alcohol dehydrogenase, acetone, when present, prevents DPNH release from the enzyme so that the mechanism at equal rates from E-DPN-2-propanol so tht the reaction is random in this direction.

References

Jan 1, 1979·Methods in Enzymology·W W Cleland
Dec 2, 1975·Biochemistry·M I SchimerlikW W Cleland
Sep 16, 1980·Biochemistry·J S Blanchard, W W Cleland
Sep 1, 1963·Biochemistry·C C WRATTEN, W W CLELAND

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Citations

Jul 1, 1993·Biotechnology and Bioengineering·J B FordK B Taylor
Jun 10, 1985·Biochimica Et Biophysica Acta·I I Nuiry, P F Cook
Apr 17, 1992·Biochimica Et Biophysica Acta·S Q LiuS K Srivastava
Apr 13, 1994·Biochimica Et Biophysica Acta·A BhatnagarS K Srivastava
Mar 17, 2005·Biochimica Et Biophysica Acta·Kurt M BohrenDavid H T Harrison
Apr 16, 1998·Biochimica Et Biophysica Acta·L E Parmentier, J S Kristensen
Dec 9, 2010·Biochemistry·Clarissa M CzeksterJohn S Blanchard
Sep 22, 2011·Biochemistry·Rafael G Silva, Vern L Schramm
Feb 2, 2008·Proceedings of the National Academy of Sciences of the United States of America·Liviu M Mirica, Judith P Klinman
May 11, 2010·The Journal of Biological Chemistry·Stefania HanauFranco Dallocchio
Oct 6, 1998·Annual Review of Biochemistry·V L Schramm
Jul 1, 1997·Isotopes in Environmental and Health Studies·P F Cook
Jun 5, 2013·Biochemistry·Alan R RendinaBenjamin Schwartz
Jan 1, 1982·CRC Critical Reviews in Biochemistry·W W Cleland
Aug 27, 2014·Biochemistry·Savarimuthu Baskar RajBryce V Plapp
Feb 6, 2008·Metabolism: Clinical and Experimental·Stephanie R AndersonStephen F Previs
Jun 1, 1991·Archives of Biochemistry and Biophysics·A BhatnagarS K Srivastava
Mar 1, 1988·Archives of Biochemistry and Biophysics·A BhatnagarS K Srivastava
Dec 21, 2006·Archives of Biochemistry and Biophysics·Isabel O FonsecaDiógenes S Santos
Aug 15, 1986·Archives of Biochemistry and Biophysics·P F FitzpatrickJ J Villafranca
Sep 2, 2000·Biophysical Journal·D B Northrop, Y K Cho
Jul 30, 2011·Archives of Biochemistry and Biophysics·Kostyantyn D BobykPaul F Cook
Jan 1, 1984·Xenobiotica; the Fate of Foreign Compounds in Biological Systems·A Y LuG T Miwa
Mar 12, 2015·Drug Metabolism and Pharmacokinetics·Zachary E TibbsCharles N Falany
May 24, 2011·Journal of the American Chemical Society·Rafael G SilvaVern L Schramm

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