Rates of phosphorylation and dephosphorylation of phosphoglycerate mutase and bisphosphoglycerate synthase

J Biol Chem. 1976 Aug 25;251(16):4817-22.

Abstract

Phosphoglycerate mutase and bisphosphoglycerate synthase (mutase) can both be phosphorylated by either glycerate-1,3-P2 or glycerate-2,3-P2 to form phosphohistidine enzymes. The present study uses a rapid quench procedure to determine if, for each enzyme, the formation of the phosphorylated enzyme and phosphate transfer from the enzyme can occur at rates consistent with the overall reactions. With bisphosphoglycerate synthase from horse red blood cells (glycerate-1,3-P2 leads to glycerate-2,3-P2) at pH 7.5, 25 degrees, phosphorylation of the enzyme appears rate-limiting, k = 13.5 s-1, compared with kcat = 12.5 s-1 for the overall synthase rate. Phosphoryl transfer from the enzyme to phosphoglycerate occurs at 38 s-1 at 4 degrees and was too fast to measure at 25 degrees. With chicken muscle phosphoglycerate mutase the half-times were too short to measure under optimal conditions. The rate of enzyme phosphorylation by glycerate-2,3-P2 at pH 5.5, 4 degrees, could account for the overall reaction rate of 170 s-1. The rate of phosphoryl transfer from the enzyme to glycerate-3-P was too rapid to measure under the same conditions. It is concluded that the phosphorylated enzymes have kinetic properties consistent with their participation as intermediates in the reactions catalyzed by these enzymes.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Bisphosphoglycerate Mutase / metabolism*
  • Chickens
  • Diphosphoglyceric Acids
  • Erythrocytes / enzymology
  • Horses
  • Hydrogen-Ion Concentration
  • Kinetics
  • Muscles / enzymology
  • Phosphoglycerate Kinase / metabolism*
  • Phosphotransferases / metabolism*
  • Protein Kinases / metabolism*
  • Time Factors

Substances

  • Diphosphoglyceric Acids
  • Phosphotransferases
  • Protein Kinases
  • Phosphoglycerate Kinase
  • Bisphosphoglycerate Mutase