title: Insulin increases branched-chain alpha-ketoacid dehydrogenase kinase expression in Clone 9 rat cells.
abstract: The branched-chain amino acids (BCAA) are committed to catabolism by the activity of the branched-chain alpha-ketoacid dehydrogenase (BCKD) complex. BCKD activity is regulated through the action of the complex-specific BCKD kinase that phosphorylates two serine residues in the E1alpha subunit. Greater BCKD kinase expression levels result in a lower activity state of BCKD and thus a decreased rate of BCAA catabolism. Activity state varies among tissues and can be altered by diet, exercise, hormones, and disease state. Within individual tissues, the concentration of BCKD kinase reflects the activity state of the BCKD complex. Here we investigated the effects of insulin, an important regulator of hepatic metabolic enzymes, on BCKD kinase expression in Clone 9 rat cells. Insulin effected a twofold increase in message levels and a twofold increase in BCKD kinase protein levels. The response was completely blocked by treatment with LY-294002 and partially blocked by rapamycin, thus demonstrating a dependence on phosphatidylinositol 3-kinase and mTOR function, respectively. These studies suggest that insulin acts to regulate BCAA catabolism through stimulation of BCKD kinase expression.
authors.affiliation: Graduate Program in Nutrition and Health Sciences, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
authors.forename: Mary M
authors.initials: MM
authors.lastname: Nellis
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Doering
authors.forename: Andrea
authors.initials: A
authors.lastname: Kasinski
authors.forename: Dean J
authors.initials: DJ
authors.lastname: Danner
mesh_terms: D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D000818:Animals; D001798:Blood Proteins / Q000494:pharmacology; D002867:Chromones / Q000494:pharmacology; D002999:Clone Cells; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D022781:Hepatocytes / Q000166:cytology / Q000201:enzymology; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D007328:Insulin / Q000494:pharmacology*; D007658:Ketone Oxidoreductases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D009025:Morpholines / Q000494:pharmacology; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats
chemical_list: D001798:Blood Proteins; D002867:Chromones; D004791:Enzyme Inhibitors; D007004:Hypoglycemic Agents; D007328:Insulin; D009025:Morpholines; D009097:Multienzyme Complexes; D012333:RNA, Messenger; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D007658:Ketone Oxidoreductases; D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D019869:Phosphatidylinositol 3-Kinases