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Articles in PresS, published online ahead of print October 16, 2001
Am J Physiol Endocrinol Metab, 10.1152/ajpendo.00101.2001
Submitted on March 7, 2001
Accepted on October 14, 2001
2 ACTIVITY AND ACETYL-CoA CARBOXYLASE PHOSPHORYLATION DURING EXERCISE
1 Physiology, Monash University, Clayton, Victoria, Australia
2 St. Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia
* To whom correspondence should be addressed. E-mail: g.mcconell{at}med.monash.edu.au.
The effect of prolonged moderate intensity exercise on human skeletal muscle AMP-activated protein kinase (AMPK)
1 and
2 activity, and acetyl-CoA carboxylase (ACCß) 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 VO2 peak with muscle biopsies obtained from the vastus lateralis at rest, 5 and 30 min of exercise. AMPK
1 activity was not altered by exercise; however, AMPK
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. ACCß 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 AMPK
2 activity were significantly correlated with both increases in ACCß 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:ATP ratio were higher (P<0.05) at both 5 min and 30 min of exercise compared with at rest. At 30 min of exercise the values of these metabolites were not significantly different to 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 AMPK
2 was associated with both nNOSµ and ACCß consistent with them both being substrates of AMPK
2 in vivo. In conclusion, AMPK
2 activity and ACCß 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.
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