CAREER: The Role of Dynamics in Enzyme Function by NMR, Chemical Synthesis, and Biochemistry

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This CAREER project addresses fundamental questions regarding the role of intramolecular protein dynamics in enzyme catalysis. Chemical synthesis of novel ligands and biophysical characterization of enzyme/ligand complexes will be combined with solution NMR spin-relaxation experiments to provide a richer understanding of the coordination between protein flexibility and protein function. These studies will be performed with the enzyme Ribonuclease A. NMR experiments will probe protein motion on distinct timescales relevant to those of biological function. Characterization of ps - ns motions will allow dissection of site-specific thermodynamic contributions involved in enzyme complex formation. Motions in the ms - ms regime are of similar timescale to chemical and physical processes essential for catalysis. Their characterization will provide atomic resolution of motions involved in a catalytically active enzyme. These experiments will provide the level of understanding of physico-chemical forces in enzyme function essential for the design of biocatalysts and optimization of protein/ligand interactions. To impact scientists and non-scientists alike, research and education must be integrated. Efforts to achieve this will focus on research mentoring, teaching, and the promotion of interaction among research groups. Incorporated within these three activities will be training and teaching of students at the undergraduate and graduate levels and from geographically underrepresented regions in an effort to simultaneously advance discovery and learning. First, the principal component of the educational plan consists of teaching two courses, 'Biochemical rates and mechanisms' and 'NMR spectroscopy.' These courses are targeted to beginning graduate students and advanced undergraduates. Both courses combine theory with experimental applications and rely on problem solving and student presentation to provide an active learning environment. Second, a major effort will be made as research mentor to graduate and undergraduate students. In addition to students from Yale, a student from rural West Virginia will be invited to perform research each summer. Finally, to promote interactions among students from diverse research groups, monthly seminars will be organized and students will be encouraged to attend and participate in discussions.