ABSTRACT
This study was designed to compare functional effects of phosphorylation of muscle acetyl-CoA carboxylase (ACC) by adenosine 3',5'-cyclic monophosphate-dependent protein kinase (PKA) and by AMP-activated protein kinase (AMPK). Muscle ACC (272 kDa) was phosphorylated and then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography. Functional effects of phosphorylation were determined by measuring ACC activity at different concentrations of each of the substrates and of citrate, an activator of the enzyme. The maximal velocity (Vmax) and the Michaelis constants (Km) for ATP, acetyl-CoA, and bicarbonate were unaffected by phosphorylation by PKA. Phosphorylation by AMPK increased the Km for ATP and acetyl-CoA. Sequential phosphorylation by PKA and AMPK, first without label and second with label, appeared to reduce the extent of label incorporation, regardless of the order. The activation constant (Ka) for citrate activation was increased to the same extent by AMPK phosphorylation, regardless of previous or subsequent phosphorylation by PKA. Thus muscle ACC can be phosphorylated by PKA but with no apparent functional effects on the enzyme. AMPK appears to be the more important regulator of muscle ACC.
Subject(s)
Acetyl-CoA Carboxylase/drug effects , Adenosine Monophosphate/pharmacology , Cyclic AMP-Dependent Protein Kinases/drug effects , Muscle, Skeletal/drug effects , Protein Kinases/drug effects , Acetyl-CoA Carboxylase/metabolism , Animals , Cyclic AMP-Dependent Protein Kinases/metabolism , Dose-Response Relationship, Drug , Muscle, Skeletal/metabolism , Phosphorylation , Protein Kinases/metabolism , RatsABSTRACT
The purpose of this study was to determine the effect of 12 wk of stairclimbing with and without an external load on aerobic capacity and quadriceps strength of sedentary (initial VO2max 25.3 +/- 0.73 ml.kg-1.min-1) (mean +/- SEM) middle aged females (50-65 yr). Three groups, LOAD (stairclimbing with external load, N = 8), STAIR (no load, N = 9), and CONTROL (N = 7) were tested. By week 4, subjects warmed up 5 min on a cycle ergometer followed by 35 min on the stairclimber at 80-85% maximum heart rate (MHR) 4 d.wk-1. In week 6, the LOAD group carried an external load of 4% of body weight increasing to 8% for weeks 7-12. STAIR and LOAD group significantly increased (P < 0.01) VO2max by 11.1% and 9.6%, respectively. Isokinetic strength tests showed increased (P < 0.05) peak torque and total work for STAIR and LOAD at 120 and 180 degrees.s-1. For total work, a significant increase (P < 0.05) of 10.5% was observed at 60 degrees.s-1 for the LOAD group. The results indicate stairclimbing is an appropriate exercise for middle-aged females improving both aerobic capacity and strength following 12 wk of training.
