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Originally published In Press as doi:10.1074/jbc.M002544200 on April 10, 2000

J. Biol. Chem., Vol. 275, Issue 30, 23146-23153, July 28, 2000
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Mechanism of Catalysis of the Cofactor-independent Phosphoglycerate Mutase from Bacillus stearothermophilus
CRYSTAL STRUCTURE OF THE COMPLEX WITH 2-PHOSPHOGLYCERATE*

Mark J. JedrzejasDagger §, Monica Chander, Peter Setlow, and Gunasekaran KrishnasamyDagger

From the Dagger  Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35294 and the  Department of Biochemistry, University of Connecticut Health Center, Farmington, Connecticut 06030

The structure of the complex between the 2,3-diphosphoglycerate-independent phosphoglycerate mutase (iPGM) from Bacillus stearothermophilus and its 3-phosphoglycerate substrate has recently been solved, and analysis of this structure allowed formulation of a mechanism for iPGM catalysis. In order to obtain further evidence for this mechanism, we have solved the structure of this iPGM complexed with 2-phosphoglycerate and two Mn2+ ions at 1.7-Å resolution. The structure consists of two different domains connected by two loops and interacting through a network of hydrogen bonds. This structure is consistent with the proposed mechanism for iPGM catalysis, with the two main steps in catalysis being a phosphatase reaction removing the phosphate from 2- or 3-phosphoglycerate, generating an enzyme-bound phosphoserine intermediate, followed by a phosphotransferase reaction as the phosphate is transferred from the enzyme back to the glycerate moiety. The structure also allowed the assignment of the function of the two domains of the enzyme, one of which participates in the phosphatase reaction and formation of the phosphoserine enzyme intermediate, with the other involved in the phosphotransferase reaction regenerating phosphoglycerate. Significant structural similarity has also been found between the active site of the iPGM domain catalyzing the phosphatase reaction and Escherichia coli alkaline phosphatase.


* This work was supported in part by a supplement to National Institutes of Health Grant GM19698 (to P. S.) for the determination of high resolution structures (to P. S. and M. J. J.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The atomic coordinates and the structure factors (code 2ejj) have been deposited in the Protein Data Bank, Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (http://www.rcsb.org/).

§ To whom correspondence should be addressed: Dept. of Microbiology, University of Alabama at Birmingham, 933 19th St. S., CHSB-19 Rm. 545, Birmingham, AL 35294-2041. Tel.: 205-975-7627; Fax: 205-975-5424; E-mail: jedrzejas@uab.edu.


Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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