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. 2003 Nov;43(1-3):97–103. doi: 10.1023/B:CYTO.0000039898.44839.90

Involvement of Protein Kinase C Activation in L-Leucine-Induced Stimulation of Protein Synthesis in L6 Myotubes

Kazumi Yagasaki 1, Naoko Morisaki 1, Yoshiro Kitahara 1, Atsuhito Miura 1, Ryuhei Funabiki 1
PMCID: PMC3449606  PMID: 19003213

Abstract

Effects of leucine and related compounds on protein synthesis were studied in L6 myotubes. The incorporation of [3H]tyrosine into cellular protein was measured as an index of protein synthesis. In leucine-depleted L6 myotubes, leucine and its keto acid, α-ketoisocaproic acid (KIC), stimulated protein synthesis, while D-leucine did not. Mepacrine, an inhibitor of both phospholipases A2 and C, canceled stimulatory actions of L-leucine and KIC on protein synthesis. Neither indomethacin, an inhibitor of cyclooxygenase, nor caffeic acid, an inhibitor of lipoxygenase, diminished their stimulatory actions, suggesting no involvement of arachidonic acid metabolism. Conversely, 1-O-hexadecyl-2-O-methylglycerol, an inhibitor of proteinkinase C, significantly canceled the stimulatory actions of L-leucine and KIC on protein synthesis, suggesting an involvement of phosphatidylinositol degradation and activation of protein kinase C. L-Leucine caused a rapid activation of protein kinase C in both cytosol and membrane fractions of the cells. These results strongly suggest that both L-leucine and KIC stimulate protein synthesis in L6 myotubes through activation of phospholipase C and protein kinase C.

Keywords: L-Leucine, L6 myotube, protein kinase C inhibitor, protein synthesis

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