Divergent Function in the Crotonase Superfamily: An Anhydride Intermediate in the Reaction Catalyzed by 3-Hydroxyisobutyryl-CoA Hydrolase
| Autor: | John A. Gerlt, Brian J. Wong |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
chemistry.chemical_classification
Binding Sites 3-hydroxyisobutyryl-CoA hydrolase Stereochemistry Chemistry Hydrolysis General Chemistry Enoyl-CoA hydratase Biochemistry Catalysis Anhydrides Enzyme catalysis Oxygen Colloid and Surface Chemistry Enzyme Pseudomonas aeruginosa Hydrolase Thiolester Hydrolases Binding site Enoyl-CoA Hydratase Function (biology) |
| Zdroj: | Journal of the American Chemical Society. 125:12076-12077 |
| ISSN: | 1520-5126 0002-7863 |
| DOI: | 10.1021/ja037652i |
| Popis: | 3-Hydroxyisobutyryl-CoA hydrolase (HICH), a member of the enoyl-CoA (crotonase) superfamily, catalyzes the hydrolysis of 3-hydroxyisobutyryl-CoA to 3-hydroxyisobutyrate. Like other members of the superfamily, the sequence of HICH contains conserved sequences for an oxyanion hole that stabilizes anionic intermediates. In contrast to most members of the superfamily, the reaction catalyzed by HICH does not proceed via formation of a thioester enolate anion; instead, evidence based on substrate deuterium isotope effects, the reactivity of substrate analogues that cannot form thioester enolate anions, single-turnover experiments in H218O, and the kinetic phenotypes of site-directed mutants provide evidence for a mechanism involving the formation of an anhydride intermediate involving Glu143 in the active site. In the reactions catalyzed by many members of the superfamily, homologues of Glu143 abstract the alpha proton of the thioester substrate to generate the thioester enolate anion intermediate. Presumably, one or more of the anionic tetrahedral intermediates on the HICH reaction coordinate are stabilized by the oxyanion hole. Thus, we conclude that the conserved oxyanion hole in this superfamily can be used to stabilize a variety of anionic intermediates. |
| Databáze: | OpenAIRE |
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