The effect of phosphate on the unfolding-refolding of phosphoglycerate kinase induced by guanidine hydrochloride.
Phosphate ions were found to stabilize the native structure of phosphoglycerate kinase without modifying the folding pathway. The transition curves obtained from different signals: enzyme activity, ellipticity at 220 nm and fluorescence intensity at 336 nm (excitation at 292 nm) are shifted to smaller guanidine hydrochloride cm values in the absence of phosphate. The kinetic characteristics are qualitatively similar, unfolding rate constants being slightly smaller in the presence of phosphate. The mechanism by which the native structure of phosphoglycerate kinase is stabilized by phosphate probably occurs upon specific phosphate binding to the nucleotide beta- or gamma-phosphate binding site of nucleotides.
Chardot
T
T
Laboratoire d'Enzymologie Physico-Chimique et Moléculaire, GR-CNRS associé à l'Université de Paris-Sud, Orsay, France.
Mitraki
A
A
Amigues
Y
Y
Desmadril
M
M
Betton
J M
JM
Yon
J M
JM
D000818:Animals; D001665:Binding Sites; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology*; D006736:Horses; D009132:Muscles / Q000201:enzymology; D010710:Phosphates / Q000494:pharmacology*; D010735:Phosphoglycerate Kinase / Q000378:metabolism*; D011489:Protein Denaturation; D013816:Thermodynamics
D006146:Guanidines; D010710:Phosphates; D010735:Phosphoglycerate Kinase; D019791:Guanidine