Properties of the phosphofructokinase regulatory factors.
The work presented herein describes many of the physiological properties of the phosphofructokinase regulatory factors. Factor activity can be separated into two discrete fractions, which were designated factor A and factor B, based on their respective charges. A preparation containing both factor A and factor B did not protect the following key carbohydrate-metabolizing enzymes from thermal inactivation: glucokinase, glucose-6-phosphatase (solubilized or nonsolubilized forms), pyruvate kinase, glucose-6-P dehydrogenase, muscle-type phosphofructokinase, or the minor liver phosphofructokinase isozyme. Factor activity in this sample was found to be Pronase sensitive, irreversibly precipitated by trichloroacetic acid, reversibly precipitated by adjusting the sample to a pH of 3.0, and stable to heating at 98 degrees C for 20 min. Distribution studies indicated that factor activity was found only in the soluble cell fraction and not in the mitochondrial or nuclear fractions. Factor activity was retained by 12,000-14,000 molecular weight cut-off (MWCO) dialysis tubing, and not retained by 50,000 MWCO dialysis tubing. These studies indicate that fructose-2,6-P2, calmodulin, or insulin-generated mediator are not associated with factor activity. Although fructose-2,6-P2 did not, both factor preparations protect the major liver phosphofructokinase isozyme (liver PFK) from inactivation by lysosomal extracts. In the diabetic rat, the activities of both factors are greatly reduced but return to near normal levels after 48 h of insulin administration. These data suggest that factor B had little or no effect on the kinetic properties of liver PFK. However, factor A was a K-type activator with respect to fructose-6-P, increasing both the Km and Ki for ATP, and slightly increasing the Vm.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D050260:Carbohydrate Metabolism; D055598:Chemical Phenomena; D002621:Chemistry; D003921:Diabetes Mellitus, Experimental / Q000201:enzymology; D005635:Fructosediphosphates / Q000378:metabolism*; D006598:Hexosediphosphates / Q000378:metabolism*; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology; D010455:Peptides / Q000378:metabolism*; D010732:Phosphofructokinase-1 / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D000596:Amino Acids; D005635:Fructosediphosphates; D006598:Hexosediphosphates; D007328:Insulin; D010455:Peptides; C043951:phosphofructokinase regulatory factors; C027652:fructose 2,6-diphosphate; D010732:Phosphofructokinase-1