The structures of complexes of yeast enolase and rabbit muscle pyruvate kinase have been solved to high resolution by X-ray crystallography. These structures reveal the identities of active-site residues in these two enzymes. Site-directed mutagenesis has been performed to clarify the roles of the active-site residues in catalysis. Residues believed to function as the acid/base catalysts for these enzymes are lysine 345 for yeast enolase and either arginine 72 or threonine 327 for pyruvate kinase. Both enzymes catalyze alternate reactions. NMR characterization of the products of these alternate reactions will be carried to help corroborate our hypotheses. NMR identification of the stereospecificity of reactions catalyzed by these enzymes and mutants will also be employed.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR002301-15
Application #
6298208
Study Section
Project Start
1999-03-01
Project End
2000-02-29
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
15
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
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