Isozyme analysis of human polymorphonuclear leukocyte phosphofructokinase from insulin resistant individuals.
Phosphofructokinase (PFK) from human polymorphonuclear leukocytes (PMN) was characterized by immunological titration with subunit specific antibodies and column chromatography on QAE-Sephadex in three different groups: control, type II diabetic, and obese individuals. It was found that PMN phosphofructokinase in the three groups consists mainly of a mixture of L4 and M4 homotetramers with possibly some hybrid forms. The predominant subunit was the L-type. A 24% decrease in the specific activity of the L-type isozyme was observed and an intermediate form (I-isozyme) having 23% of the total activity in diabetic individuals appeared. In obese individuals a 30% decrease was observed in the activity of M-type isozyme and 9% of the total activity corresponded to the intermediate form. Kinetic studies showed different regulatory properties among the isozymes from the three groups. The lower PFK activity found in diabetic and obese individuals can be associated with the decreased activity in the L-type isozyme (for diabetic individuals) and in the M-type isozyme (for obese individuals); the lower activity can also be associated with the four times lower affinity for F-6-P showed by the M-type isozyme, the decreased sensitivity to ATP inhibition (for both isozymes), and the appearance of an intermediate form with a different kinetic behaviour.
Durante
P
P
Instituto de Investigaciones Clínicas, Facultad de Medicina, Universidad del Zulia, Maracaibo, Venezuela.
Raleigh
X
X
Gómez
M E
ME
Campos
G
G
Ryder
E
E
D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D016022:Case-Control Studies; D002951:Citrates / Q000494:pharmacology; D019343:Citric Acid; D003924:Diabetes Mellitus, Type 2 / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005635:Fructosediphosphates / Q000494:pharmacology; D005636:Fructosephosphates / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007333:Insulin Resistance / Q000502:physiology*; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000097:blood*; D007700:Kinetics; D009504:Neutrophils / Q000201:enzymology*; D009765:Obesity / Q000201:enzymology; D010732:Phosphofructokinase-1 / Q000037:antagonists & inhibitors / Q000097:blood*; D013379:Substrate Specificity
D002951:Citrates; D004791:Enzyme Inhibitors; D005635:Fructosediphosphates; D005636:Fructosephosphates; D007527:Isoenzymes; D019343:Citric Acid; C027618:fructose-6-phosphate; D000255:Adenosine Triphosphate; D010732:Phosphofructokinase-1; C029063:fructose-1,6-diphosphate