CAREER: Determination of enzyme mechanisms using new kinetic isotope effect approaches

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With this CAREER award, the Chemistry of Life Processes Program is supporting Dr. Andrew Murkin of the University at Buffalo, The State University of New York, in the development of new methods for studying the mechanisms of enzymatic reactions. The objective of this research is to determine the transition state structures of two enzymes that catalyze reactions with unresolved mechanisms. To reach this goal, Dr. Murkin developed a method for measuring kinetic isotope effects based on two-dimentional NMR spectroscopy that delivers better sensitivity and precision than previous methods. Transition state models will be constructed by matching the theoretical kinetic isotope effects obtained computationally to those measured experimentally.When a molecule undergoes a reaction and reaches the transition state, the atoms that react or experience geometric or electrostatic changes exert differences in rates when substituted with heavier isotopes. Such kinetic isotope effects provide an experimentally accessible picture of the substrate during the rate-limiting step in a mechanism. The techniques developed from this award should be of interest to a wide audience of scientists who depend on kinetic isotope effects to study a variety of processes, including the design of new therapies. This program is also designed to encourage youth to pursue training in biochemistry and related sciences. To stimulate educational interest among youth, an introduction to enzymes will be provided to K-12 students in greater-Buffalo schools, including underrepresented youths, through informative hands-on, inquiry-based experiments conducted in middle schools and in a university laboratory. At the university level, the design of transition state inhibitors and measurement of kinetic isotope effects will be integrated into undergraduate and graduate chemistry courses.