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Near attack conformers dominate β-phosphoglucomutase complexes where geometry and charge distribution reflect those of substrate
Joanna L. Griffin, Matthew W. Bowler, Nicola J. Baxter, Katherine N. Leigh, Hugh R. W. Dannatt, Andrea M. Hounslow, G. Michael Blackburn, Charles Edwin Webster, Matthew J. Cliff and Jonathan P. Waltho
Proceedings of the National Academy of Sciences of the United States of America
Vol. 109, No. 18 (May 1, 2012), pp. 6910-6915
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/41596104
Page Count: 6
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Experimental observations of fluoromagnesate and fluoroaluminate complexes of β-phosphoglucomutase (β-PGM) have demonstrated the importance of charge balance in transition-state stabilization for phosphoryl transfer enzymes. Here, direct observations of ground-state analog complexes of β-PGM involving trifluoroberyllate establish that when the geometry and charge distribution closely match those of the substrate, the distribution of conformers in solution and in the crystal predominantly places the reacting centers in van der Waals proximity. Importantly, two variants are found, both of which satisfy the criteria for near attack conformers. In one variant, the aspartate general base for the reaction is remote from the nucleophile. The nucleophile remains protonated and forms a nonproductive hydrogen bond to the phosphate surrogate. In the other variant, the general base forms a hydrogen bond to the nucleophile that is now correctly orientated for the chemical transfer step. By contrast, in the absence of substrate, the solvent surrounding the phosphate surrogate is arranged to disfavor nudeophilic attack by water. Taken together, the trifluoroberyllate complexes of β-PGM provide a picture of how the enzyme is able to organize itself for the chemical step in catalysis through the population of intermediates that respond to increasing proximity of the nucleophile. These experimental observations show how the enzyme is capable of stabilizing the reaction pathway toward the transition state and also of minimizing unproductive catalysis of aspartyl phosphate hydrolysis.
Proceedings of the National Academy of Sciences of the United States of America © 2012 National Academy of Sciences