Analysis of the phosphofructokinase subunits and isoenzymes in human tissues.
The 6-phosphofructo-1-kinase (PFK) subunits and isoenzymes were studied in human muscle, heart, brain, liver, platelets, fibroblasts, erythrocytes, placenta and umbilical cord. In each tissue, the subunit types in the native isoenzymes were characterized by immunological titration with subunit-specific antibodies and by column chromatography on QAE (quaternary aminoethyl)-Sephadex. Further, the subunits of the partially purified native isoenzymes were resolved by SDS/polyacrylamide-gel electrophoresis, identified by immunoblotting, and quantified by scanning gel densitometry of silver-stained gels and immunoblots. Depending on the type of tissue, one to three subunits were detected. The Mr values of the L, M and C subunits regardless of tissue were 76,700 +/- 1400, 82,500 +/- 1640 and 86,500 +/- 1620. Of the tissues studied, only the muscle PFK isoenzymes exhibited one subunit, which was the M-type subunit. Of the other tissues studied, the PFK isoenzymes contained various amounts of all three subunits. Considering the properties of the native PFK isoenzymes, it is clear that, in human tissues, they are not simply various combinations of two or three homotetrameric isoenzymes, but complex mixtures of homotetramers and heterotetramers. The kinetic/regulatory properties of the various isoenzyme pools were found to be dependent on subunit composition.
Dunaway
G A
GA
Department of Pharmacology, Southern Illinois University School of Medicine, Springfield 62794-9230.
Kasten
T P
TP
Sebo
T
T
Trapp
R
R
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000293:Adolescent; D004591:Electrophoresis, Polyacrylamide Gel; D005636:Fructosephosphates / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D008297:Male; D010732:Phosphofructokinase-1 / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D014018:Tissue Distribution
D005636:Fructosephosphates; D007074:Immunoglobulin G; D007527:Isoenzymes; C027618:fructose-6-phosphate; D000255:Adenosine Triphosphate; D010732:Phosphofructokinase-1