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Originally published in Science Express on 13 March 2003
Science 28 March 2003:
Vol. 299. no. 5615, pp. 2067 - 2071
DOI: 10.1126/science.1082710
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Reports

The Pentacovalent Phosphorus Intermediate of a Phosphoryl Transfer Reaction

Sushmita D. Lahiri,1 Guofeng Zhang,2 Debra Dunaway-Mariano,2* Karen N. Allen1*

Enzymes provide enormous rate enhancements, unmatched by any other type of catalyst. The stabilization of high-energy states along the reaction coordinate is the crux of the catalytic power of enzymes. We report the atomic-resolution structure of a high-energy reaction intermediate stabilized in the active site of an enzyme. Crystallization of phosphorylated beta -phosphoglucomutase in the presence of the Mg(II) cofactor and either of the substrates glucose 1-phosphate or glucose 6-phosphate produced crystals of the enzyme-Mg(II)-glucose 1,6-(bis)phosphate complex, which diffracted x-rays to 1.2 and 1.4 angstroms, respectively. The structure reveals a stabilized pentacovalent phosphorane formed in the phosphoryl transfer from the C(1)O of glucose 1,6-(bis)phosphate to the nucleophilic Asp8 carboxylate.

1 Department of Physiology and Biophysics, Boston University School of Medicine, Boston, MA 02118-2394, USA.
2 Department of Chemistry, University of New Mexico, Albuquerque, NM 87131, USA.
*   To whom correspondence should be addressed. E-mail: allen{at}med-xtal.bu.edu (K.N.A.); dd39{at}unm.edu (D.D.-M.)

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