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Am J Physiol Endocrinol Metab 282: E688-E694, 2002. First published October 16, 2001; doi:10.1152/ajpendo.00101.2001
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Vol. 282, Issue 3, E688-E694, March 2002

Progressive increase in human skeletal muscle AMPKalpha 2 activity and ACC phosphorylation during exercise

T. J. Stephens1,*, Z.-P. Chen2,*, B. J. Canny1, B. J. Michell2, B. E. Kemp2, and G. K. McConell1

1 Department of Physiology, Monash University, Clayton, Victoria 3800; and 2 St. Vincent's Institute of Medical Research, Fitzroy, Victoria 3065, Australia

The effect of prolonged moderate-intensity exercise on human skeletal muscle AMP-activated protein kinase (AMPK)alpha 1 and -alpha 2 activity and acetyl-CoA carboxylase (ACCbeta ) and neuronal nitric oxide synthase (nNOSµ) phosphorylation was investigated. Seven active healthy individuals cycled for 30 min at a workload requiring 62.8 ± 1.3% of peak O2 consumption (VO2 peak) with muscle biopsies obtained from the vastus lateralis at rest and at 5 and 30 min of exercise. AMPKalpha 1 activity was not altered by exercise; however, AMPKalpha 2 activity was significantly (P < 0.05) elevated after 5 min (~2-fold), and further elevated (P < 0.05) after 30 min (~3-fold) of exercise. ACCbeta phosphorylation was increased (P < 0.05) after 5 min (~18-fold compared with rest) and increased (P < 0.05) further after 30 min of exercise (~36-fold compared with rest). Increases in AMPKalpha 2 activity were significantly correlated with both increases in ACCbeta phosphorylation and reductions in muscle glycogen content. Fat oxidation tended (P = 0.058) to increase progressively during exercise. Muscle creatine phosphate was lower (P < 0.05), and muscle creatine, calculated free AMP, and free AMP-to-ATP ratio were higher (P < 0.05) at both 5 and 30 min of exercise compared with those at rest. At 30 min of exercise, the values of these metabolites were not significantly different from those at 5 min of exercise. Phosphorylation of nNOSµ was variable, and despite the mean doubling with exercise, statistically significance was not achieved (P = 0.304). Western blots indicated that AMPKalpha 2 was associated with both nNOSµ and ACCbeta consistent with them both being substrates of AMPKalpha 2 in vivo. In conclusion, AMPKalpha 2 activity and ACCbeta phosphorylation increase progressively during moderate exercise at ~60% of VO2 peak in humans, with these responses more closely coupled to muscle glycogen content than muscle AMP/ATP ratio.

adenosine monophosphate-activated protein kinase; acetyl-coenzyme A carboxylase-beta ; neuronal nitric oxide synthase; prolonged exercise; humans

* T. J. Stephens and Z.-P. Chen contributed equally to this study.




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