The reaction of phosphohexomutase from Pseudomonas aeruginosa: structural insights into a simple processive enzyme

J Biol Chem. 2006 Jun 2;281(22):15564-71. doi: 10.1074/jbc.M600590200. Epub 2006 Apr 4.

Abstract

The enzyme phosphomannomutase/phosphoglucomutase (PMM/PGM) from Pseudomonas aeruginosa catalyzes the reversible conversion of 1-phospho to 6-phospho-sugars. The reaction entails two phosphoryl transfers, with an intervening 180 degrees reorientation of the reaction intermediate (e.g. glucose 1,6-bisphosphate) during catalysis. Reorientation of the intermediate occurs without dissociation from the active site of the enzyme and is, thus, a simple example of processivity, as defined by multiple rounds of catalysis without release of substrate. Structural characterization of two PMM/PGM-intermediate complexes with glucose 1,6-bisphosphate provides new insights into the reaction catalyzed by the enzyme, including the reorientation of the intermediate. Kinetic analyses of site-directed mutants prompted by the structural studies reveal active site residues critical for maintaining association with glucose 1,6-bisphosphate during its unique dynamic reorientation in the active site of PMM/PGM.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Catalysis
  • Catalytic Domain / genetics
  • Crystallography, X-Ray
  • Glucose-6-Phosphate / analogs & derivatives
  • Glucose-6-Phosphate / metabolism
  • Hydrogen Bonding
  • Kinetics
  • Models, Biological
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Phosphoglucomutase / chemistry*
  • Phosphoglucomutase / genetics
  • Phosphoglucomutase / metabolism*
  • Phosphotransferases (Phosphomutases) / chemistry*
  • Phosphotransferases (Phosphomutases) / genetics
  • Phosphotransferases (Phosphomutases) / metabolism*
  • Protein Conformation
  • Pseudomonas aeruginosa / enzymology*
  • Pseudomonas aeruginosa / genetics
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Static Electricity
  • Substrate Specificity

Substances

  • Recombinant Proteins
  • Glucose-6-Phosphate
  • glucose-1,6-bisphosphate
  • Phosphotransferases (Phosphomutases)
  • Phosphoglucomutase
  • phosphomannomutase

Associated data

  • PDB/2FKF
  • PDB/2FKM