Effects of insulin and protein synthesis inhibitors on phospholipid metabolism, diacylglycerol levels, and pyruvate dehydrogenase activity in BC3H-1 cultured myocytes

R. V. Farese, D. E. Barnes, J. S. Davis, M. L. Standaert, R. J. Pollet

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Abstract

BC3H-1 myocytes were cultured with 32PO4 for 3 days to label phospholipids to constant specific activity. Subsequent treatment with physiological concentrations of insulin provoked 40-70% increases in 32PO4 levels (reflecting increases in mass) in phosphatidic acid, phosphatidylinositol, and polyphosphoinositides, and, lesser, 20-25% increases in phosphatidylserine and the combined chromatographic area containing phosphatidylethanolamine plus phosphatidylcholine plus phosphatidylcholine. Insulin-induced increases in phospholipids were significant within 5 min and near-maximal at 15-30 min. Comparable rapid insulin-induced increases in [3H]phosphatidylinositol were observed in myocytes prelabeled with [3H]inositol. These insulin effects (as per prolonged pulse-chase experiments) were due to increased phospholipid synthesis rather than decreased phospholipid degradation. Cycloheximide (and puromycin) pretreatment prevented insulin-induced increases in phospholipids and rapidly reversed ongoing insulin effects on phospholipids and pyruvate dehydrogenase activity. Insulin also rapidly increased diacylglycerol levels. These findings suggest that: (a) insulin provokes rapid increases in de novo synthesis of phosphatidic acid and its derivatives, e.g. phosphoinositides and diacylglycerol; (b) protein synthesis inhibitors diminish phospholipid levels in insulin-treated (but not control) tissues by increasing phospholipid degradation (?phospholipase(s) activation); and (c) changes in phospholipids and diacylglycerol may be important for changes in pyruvate dehydrogenase and other enzymatic activities during treatment with insulin and/or protein synthesis inhibitors.

Original languageEnglish (US)
Pages (from-to)7094-7100
Number of pages7
JournalJournal of Biological Chemistry
Volume259
Issue number11
StatePublished - Aug 23 1984

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Protein Synthesis Inhibitors
Diglycerides
Pyruvic Acid
Metabolism
Muscle Cells
Phospholipids
Oxidoreductases
Insulin
Phosphatidylinositols
Phosphatidic Acids
Phosphatidylcholines
Phosphatidylinositol Phosphates
Degradation
Puromycin
Phospholipases
Phosphatidylserines
Inositol
Cycloheximide
Labels
Chemical activation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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Effects of insulin and protein synthesis inhibitors on phospholipid metabolism, diacylglycerol levels, and pyruvate dehydrogenase activity in BC3H-1 cultured myocytes. / Farese, R. V.; Barnes, D. E.; Davis, J. S.; Standaert, M. L.; Pollet, R. J.

In: Journal of Biological Chemistry, Vol. 259, No. 11, 23.08.1984, p. 7094-7100.

Research output: Contribution to journalArticle

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