Phosphofructokinase binding to myofibrils in fish muscle: influences of ionic strength and metabolite levels on enzyme complex formation.
The interaction of 6-phosphofructo-1-kinase (PFK) with myofibrils was assessed using the purified proteins from rainbow trout white skeletal muscle. More than 70% of PFK activity was bound at pH values between 6 and 7 but higher pHs dissociated the complex. Increasing salt concentrations also reduced PFK binding, with greater sensitivity to salt at pH 7.0 vs pH 6.6. Substrates and allosteric effectors also reduced enzyme binding to myofibrils; 50% of enzyme was released at 0.23, 0.24, 1.3, and 2.0 mM for fructose 6-phosphate, Mg.ATP, ATP and AMP, respectively. However, the addition of a protein crowding agent, poly(ethylene)glycol, greatly enhanced PFK binding to myofibrils, particularly at high pH (8.0) or high [KCl]. The studies suggest that reversible binding of PFK to myofibrils may be an important factor in the control of anaerobic glycolysis in vivo, especially under the cellular conditions of burst swimming exercise.
Su
J Y
JY
Institute of Biochemistry, Carleton University, Ottawa, Ontario, Canada.
Storey
K B
KB
D000199:Actins / Q000494:pharmacology; D000249:Adenosine Monophosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001667:Binding, Competitive; D002712:Chlorides / Q000494:pharmacology; D006019:Glycolysis / Q000187:drug effects*; D006863:Hydrogen-Ion Concentration; D018482:Muscle, Skeletal / Q000378:metabolism*; D009210:Myofibrils / Q000378:metabolism*; D017686:Oncorhynchus mykiss; D009994:Osmolar Concentration; D010710:Phosphates / Q000494:pharmacology; D010732:Phosphofructokinase-1 / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D017680:Potassium Compounds / Q000494:pharmacology
D000199:Actins; D002712:Chlorides; D010710:Phosphates; D017680:Potassium Compounds; D011092:Polyethylene Glycols; D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate; C013216:potassium phosphate; D010732:Phosphofructokinase-1