Effects of diabetes on the activity and content of the branched-chain alpha-ketoacid dehydrogenase complex in liver.
Severe ketotic diabetes induced in rats by streptozotocin resulted in a reduction in activity of the hepatic branched-chain alpha-ketoacid dehydrogenase complex, regardless of whether activity was expressed on the basis of liver wet weight, total liver, liver protein, or liver DNA. A decrease in enzyme specific activity (units of enzyme activity per mg of enzyme protein) was found responsible for the reduction in measurable enzyme activity of the complex. Insulin treatment reversed the decrease in enzyme specific activity. Treatment of tissue extracts with phosphoprotein phosphatase had no effect, indicating that activity of the complex was decreased by some mechanism other than reversible phosphorylation. Specific protein components of the complex were also not found reduced by the diabetic state. Induction of severe ketotic diabetes in rats previously fed a low-protein diet resulted in activation of the enzyme as a consequence of dephosphorylation. Nevertheless, the specific activity of the dephosphorylated enzyme of diabetic, low-protein-fed rats was decreased relative to that of control, low-protein-fed animals. Reconstitution studies with tissue extracts fortified with the purified E1 component indicate that severe diabetes induces a defect in this component of the hepatic branched-chain alpha-ketoacid dehydrogenase complex.
Gibson
R
R
Department of Biochemistry, Indiana University School of Medicine, Indianapolis 46202-5122.
Zhao
Y
Y
Jaskiewicz
J
J
Fineberg
S E
SE
Harris
R A
RA
D020155:3-Hydroxybutyric Acid; D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D000090:Acetoacetates / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002950:Citrate (si)-Synthase / Q000378:metabolism; D003921:Diabetes Mellitus, Experimental / Q000201:enzymology* / Q000378:metabolism; D006885:Hydroxybutyrates / Q000378:metabolism; D007658:Ketone Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008930:Mitochondria, Liver / Q000201:enzymology*; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D012016:Reference Values; D013311:Streptozocin / Q000494:pharmacology
D000090:Acetoacetates; D006885:Hydroxybutyrates; D009097:Multienzyme Complexes; C016635:acetoacetic acid; D013311:Streptozocin; D007658:Ketone Oxidoreductases; D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D002950:Citrate (si)-Synthase; D020155:3-Hydroxybutyric Acid