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1 Department of Human Biology and Nutritional Sciences, University of Guelph, Guelph, Ontario N1G 2W1; 2 Department of Medicine, McMaster University, Hamilton, Ontario, Canada L8N 3Z5; and 3 Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana 46202
The increase in skeletal
muscle pyruvate dehydrogenase kinase (PDK) activity was measured in
skeletal muscle of six healthy males after a eucaloric
high-fat/low-carbohydrate (HF/LC; 5% carbohydrate, 73% fat, and 22%
protein of total energy intake) diet compared with a standardized
prediet (50% carbohdyrate, 30% fat, and 21% protein). Biopsies were
obtained from the vastus lateralis muscle after 3 days on the prediet
(day 0) and after 1, 2, and 3 days of the HF/LC diet. Intact
mitchondria were extracted from fresh muscle and analyzed for PDK
activity and Western blotting of PDK2 and PDK4 protein. A second biopsy
was taken at each time point and frozen for Northern blot analysis of
PDK2 and PDK4 mRNAs. PDK activity increased in a linear fashion over
the 3-day HF/LC diet and was significantly higher than control by 1 day. PDK activity was 0.09 ± 0.03, 0.18 ± 0.05, 0.30 ± 0.07, and 0.37 ± 0.09 min
1 at 0, 1, 2, and 3 days, respectively. PDK4 protein and mRNA increased maximally by
day 1, and PDK2 protein and mRNA were unaffected by the
HF/LC diet. Resting respiratory exchange ratios decreased after 1 day
of the HF/LC diet (from 0.79 ± 0.02 to 0.72 ± 0.02) and
remained depressed throughout the 3-day dietary intervention (0.68 ± 0.01). The immediate shift to fat utilization was accompanied by
increased blood glycerol, 
-hydroxybutyrate, and plasma free fatty
acid concentrations. These results suggest that the continuing increase
in PDK activity over the 3-day HF/LC diet is not due to increasing PDK
protein beyond 1 day. This could be due to the contribution of another
isoform to the total PDK activity or to a continual increase in PDK4 or
PDK2 specific activity.
pyruvate dehydrogenase kinase 2; pyruvate dehydrogenase kinase 4; respiratory exchange ratio; pyruvate dehydrogenase activity; mitochondria
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