PMID: 7016113Nov 15, 1980Paper

Studies on the incorporation of [32P]phosphate into pyruvate dehydrogenase in intact rat fat-cells. Effects of insulin

The Biochemical Journal
W A HughesR M Denton

Abstract

1. Intact rat epididymal fat-cells were incubated with 32Pi, and the intracellular proteins were separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. One of the separated bands of phosphorylated proteins had an apparent subunit mol.wt. of 42 000, which is the same as that of the alpha-subunit of the pyruvate dehydrogenase complex. By using a combination of subcellular fractionation, immunoprecipitation with antiserum raised against pyruvate dehydrogenase complex and two-dimensional electrophoresis it was apparent that the incorporation into alpha-subunits accounted for 35--45% of the total incorporation into this band of phosphoproteins. 2. The increase in the initial activity of pyruvate dehydrogenase that follows brief exposure of fat-cells to insulin was shown to be associated with a decrease in the steady-state incorporation of 32P into the alpha-subunits of pyruvate dehydrogenase. 3. Tryptic peptide analysis of pyruvate dehydrogenase [32P]phosphate, labelled in intact fat-cells, indicated that three serine residues on the alpha-subunit were phosphorylated, corresponding to the three sites phosphorylated when purified pig heart pyruvate dehydrogenase was incubated with [gamma-32P]ATP. The relative phosph...Continue Reading

Citations

Jan 1, 1985·Diabetes/metabolism Reviews·W K Gottschalk, L Jarett
Jul 5, 1983·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·R M Denton, R W Brownsey
Jan 1, 1989·Annals of the New York Academy of Sciences·R M DentonJ G McCormack
Mar 1, 1984·Molecular and Cellular Endocrinology·B Asgeirsson, J G Schofield
Jan 1, 1982·Progress in Lipid Research·J G McCormack
Aug 1, 1987·Archives of Biochemistry and Biophysics·K Cheng, J Larner
Dec 28, 1983·Biochemical and Biophysical Research Communications·S Ramakrishna, W B Benjamin

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