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Structural and mechanistic studies of enolase
G H Reed1, R R Poyner, T M Larsen
1Institute for Enzyme Research, Graduate School, University of Wisconsin-Madison 53705, USA.
Current Opinion in Structural Biology
|December 1, 1996
Summary
Yeast enolase structure reveals how it binds substrates and stabilizes the enolate intermediate. Site-specific mutations confirm the roles of amino acid side chains in catalysis.
Area of Science:
- Biochemistry
- Enzymology
- Structural Biology
Background:
- Yeast enolase catalyzes a crucial step in glycolysis.
- Understanding enzyme mechanisms is key to metabolic pathway research.
Purpose of the Study:
- To elucidate the catalytic mechanism of yeast enolase.
- To identify key amino acid residues and metal ion roles in catalysis.
Main Methods:
- X-ray crystallography of yeast enolase with substrate/product.
- Site-directed mutagenesis and enzyme characterization.
Main Results:
- High-resolution structure reveals substrate/product binding stereochemistry.
- Identified amino acid side chains critical for acid/base catalysis.
- Magnesium ion coordination to substrate/product stabilizes the enolate intermediate.
Conclusions:
- Detailed mechanism of yeast enolase catalysis elucidated.
- Structural and mutational data confirm catalytic roles of specific residues and metal ions.
- Provides insights into enolate intermediate stabilization.