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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
Mechanism of Catalysis of the Cofactor-independent
Phosphoglycerate Mutase from Bacillus
stearothermophilus
CRYSTAL STRUCTURE OF THE COMPLEX WITH 2-PHOSPHOGLYCERATE*
Mark J.
Jedrzejas §,
Monica
Chander¶,
Peter
Setlow¶, and
Gunasekaran
Krishnasamy
From the 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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