Phosphofructokinase from bumblebee flight muscle. Molecular and catalytic properties and role of the enzyme in regulation of the fructose 6-phosphate/fructose 1,6-bisphosphate cycle.
Phosphofructokinase from the flight muscle of bumblebee was purified to homogeneity and its molecular and catalytic properties are presented. The kinetic behavior studies at pH 8.0 are consistent with random or compulsory-order ternary complex. At pH 7.4 the enzyme displays regulatory behavior with respect to both substrates, cooperativity toward fructose 6-phosphate, and inhibition by high concentration of ATP. Determinations of glycolytic intermediates in the flight muscle of insects exposed to low and normal temperatures showed statistically significant increases in the concentrations of AMP, fructose 2,6-bisphosphate, and glucose 6-phosphate during flight at 25 degrees C or rest at 5 degrees C. Measuring the activity of phosphofructokinase and fructose 1,6-bisphosphatase at 25 and 7.5 degrees C, in the presence of physiological concentrations of substrates and key effectors found in the muscle of bumblebee kept under different environmental temperatures and activity levels, suggests that the temperature dependence of fructose 6-phosphate/fructose 1,6-bisphosphate cycling may be regulated by fluctuation of fructose 2,6-bisphosphate concentration and changes in the affinity of both enzymes for substrates and effectors. Moreover, in the presence of in vivo concentrations of substrates, phosphofructokinase is inactive in the absence of fructose 2,6-bisphosphate.
Leite
A
A
Department of Biochemistry, University of São Paulo, Brazil.
Neto
J A
JA
Leyton
J F
JF
Crivellaro
O
O
el-Dorry
H A
HA
D000818:Animals; D001516:Bees / Q000201:enzymology*; D005426:Flight, Animal; D005635:Fructosediphosphates / Q000378:metabolism*; D005636:Fructosephosphates / Q000378:metabolism*; D006598:Hexosediphosphates / Q000378:metabolism*; D006706:Homeostasis; D007700:Kinetics; D009132:Muscles / Q000201:enzymology*; D010732:Phosphofructokinase-1 / Q000302:isolation & purification / Q000378:metabolism*; D013816:Thermodynamics
D005635:Fructosediphosphates; D005636:Fructosephosphates; D006598:Hexosediphosphates; C027618:fructose-6-phosphate; D010732:Phosphofructokinase-1; C029063:fructose-1,6-diphosphate