Measurement and Analysis of Kinetic Isotope Effects

Experimental Analysis of Enzyme Mechanism Using Isotope Effects, Volume 596, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Chapters in this comprehensive update include Measurement of enzyme binding isotope effects, Chemical ligation and isotope labeling to locate dynamic effects, Measurement of heavy enzyme isotope effects, Extracting kinetic isotope effects from a global analysis of reaction progress curves, KIE of metabolic flux and enzymes, Solvent and Primary KIE on Flavin Enzymes, and The Rapid Determination of Primary Deuterium Isotope Effects on Enzyme-Catalyzed Proton Transfer at Carbon in 50/50 HOH/DOD.

Readers who are interested in applying or understanding this research will find useful methods currently used for measuring isotope effects on solution and enzyme reactions.

1. Measurement of enzyme binding isotope effects Vern Schramm 2. Chemical ligation and isotope labeling to locate dynamic effects Rudolf Alleman 3. Measurement of heavy enzyme isotope effects Amnon Kohen 4. Extracting kinetic isotope effects from a global analysis of reaction progress curves Sam Hay 5. KIE of metabolic flux and enzymes Guillaume Tcherkez 6. Solvent and Primary KIE on Flavin Enzymes Paul Fitzpatrick 7. The Rapid Determination of Primary Deuterium Isotope Effects on Enzyme-Catalyzed Proton Transfer at Carbon in 50/50 HOH/DOD John Richard 8. 18-O isotope effects on oxygenases Judith Klinman 9. Chlorine isotope effects on dehalogenases Piotr Paneth 10. Chlorine isotope effects (w/Paneth) Agnieszka Dybala-Defratyka 11. Kinetic Deuterium Isotope Effects in Cytochrome P450 Reactions F. Peter Guengerich 12. Use of Isotopes and Isotope Effects for Investigations of Diiron Oxygenase Mechanisms John Lipscomb 13. Characterization of substrate, co-substrate, and product isotope effects associated with enzymatic oxygenations using gas- and liquid chromatography/isotope ratio mass spectrometry Thomas Hofstetter 14. NMR titrations to determine equilibrium isotope effects Charles Perrin 15. NMR to characterize the nonstatistical distribution of stable isotopes in metabolism Richard Robins 16. Applications of isotope ratio mass spectrometry in sports drug testing: accounting for isotope fractionation in analysis of biological samples Thomas Piper 17. Whole molecule mass spectrometry to measure KIEs on RNA 2’O-transphosphorylation reactions Mike Harris 18. Isotope Effects on the Concentration of Steady State Intermediates Hung-Wen Liu and Mark Ruszczycky 19. Headspace analysis of CO2 to measure carbon kinetic isotope effects in decarboxylation reactions Ron Kluger 20. 32P/33P remote label methods to measure RNA kinetic isotope effects Joseph Piccirilli 21. Isotope effects and MS analysis of pseudouridine synthase enzymes Eugene Mueller 22. Nucleophile isotope effects on glycosyl transfer reactions Andrew James Bennet

Enzymologists and biochemists interested in learning how to overcome the inherent challenges to measuring and interpreting kinetic isotope effects.

• Written by pioneers of modern isotope effect research
• Is the only collection of modern kinetic isotope effect methods currently available