title: Near attack conformers dominate β-phosphoglucomutase complexes where geometry and charge distribution reflect those of substrate.
abstract: 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 nucleophilic 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.
authors.affiliation: Department of Molecular Biology and Biotechnology, Krebs Institute,  University of Sheffield, Sheffield S10 2TN,  United Kingdom.
authors.forename: Joanna L
authors.initials: JL
authors.lastname: Griffin
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Bowler
authors.forename: Nicola J
authors.initials: NJ
authors.lastname: Baxter
authors.forename: Katherine N
authors.initials: KN
authors.lastname: Leigh
authors.forename: Hugh R W
authors.initials: HR
authors.lastname: Dannatt
authors.forename: Andrea M
authors.initials: AM
authors.lastname: Hounslow
authors.forename: G Michael
authors.initials: GM
authors.lastname: Blackburn
authors.forename: Charles Edwin
authors.initials: CE
authors.lastname: Webster
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Cliff
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Waltho
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D001608:Beryllium / Q000737:chemistry; D018360:Crystallography, X-Ray; D005459:Fluorides / Q000737:chemistry; D013294:Lactococcus lactis / Q000201:enzymology; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D017875:Phosphotransferases (Phosphomutases) / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D055672:Static Electricity; D013816:Thermodynamics
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; C035982:beryllium fluoride; D017875:Phosphotransferases (Phosphomutases); C512509:beta-phosphoglucomutase, Lactococcus lactis; D001608:Beryllium; D005459:Fluorides