The cooperative binding of fructose-1,6-bisphosphate to yeast pyruvate kinase.
The cooperative binding of the allosteric activator fructose-1,6-bisphosphate [Fru(1,6)P2] to yeast pyruvate kinase was investigated by equilibrium dialysis and fluorescence quench titration. The results show that yeast pyruvate kinase binds four molecules of Fru(1,6)P2 per tetramer and the observed fluorescence quench follows the binding of the ligand and not the cooperative T to R state transition. Additionally it is shown that the binding of Fru(1,6)P2 to yeast pyruvate kinase is compatible with the model of cooperativity that has been proposed and incorporates an intermediate state, R', with properties between those of the T and R states.
Murcott
T H
TH
Department of Biochemistry, School of Medical Sciences, University of Bristol, UK.
Gutfreund
H
H
Muirhead
H
H
D000494:Allosteric Regulation; D000495:Allosteric Site; D003956:Dialysis; D005635:Fructosediphosphates / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D011770:Pyruvate Kinase / Q000378:metabolism*; D012044:Regression Analysis; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013050:Spectrometry, Fluorescence
D005635:Fructosediphosphates; D011770:Pyruvate Kinase; C029063:fructose-1,6-diphosphate