XRAY ABSORPTION SPECTROSCOPY OF NUCLEOPHILIC ZN ENZYMES

$143,176P41FY2001RRNIH

Stanford University, Stanford CA

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Abstract

Recent work has revealed the existence of a novel class of Zn enzymes, in which the Zn appears to function to activate a coordinated thiolate toward nucleophilic attack. Experiments are proposed on three such enzymes that are involved in various aspects of methyl transfer: cobalamin-dependent and cobalamin-independent methionine synthase and cobamide: coenzyme M methyl transferase. EXAFS measurements on the native enzyme, on site directed mutants, and on enzyme samples with a variety of substrates and substrate analogs are proposed, with the goal of elucidating the three dimensional geometry of the Zn active sites.

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