Lysine suppresses protein degradation through autophagic-lysosomal system in C2C12 myotubes

Mol Cell Biochem. 2014 Jun;391(1-2):37-46. doi: 10.1007/s11010-014-1984-8. Epub 2014 Feb 15.


Muscle mass is determined between protein synthesis and protein degradation. Reduction of muscle mass leads to bedridden condition and attenuation of resistance to diseases. Moreover, bedridden condition leads to additional muscle loss due to disuse muscle atrophy. In our previous study (Sato et al. 2013), we showed that administered lysine (Lys), one of essential amino acid, suppressed protein degradation in skeletal muscle. In this study, we investigated that the mechanism of the suppressive effects of Lys on skeletal muscle proteolysis in C2C12 cell line. C2C12 myotubes were incubated in the serum-free medium containing 10 mM Lys or 20 mM Lys, and myofibrillar protein degradation was determined by the rates of 3-methylhistidine (MeHis) release from the cells. The mammalian target of rapamycin (mTOR) activity from the phosphorylation levels of p70-ribosormal protein S6 kinase 1 and eIF4E-binding protein 1 and the autophagic-lysosomal system activity from the ratio of LC3-II/I in C2C12 myotubes stimulated by 10 mM Lys for 0-3 h were measured. The rates of MeHis release were markedly reduced by addition of Lys. The autophagic-lysosomal system activity was inhibited upon 30 min of Lys supplementation. The activity of mTOR was significantly increased upon 30 min of Lys supplementation. The suppressive effect of Lys on the proteolysis by the autophagic-lysosomal system was maintained partially when mTOR activity was inhibited by 100 nM rapamycin, suggesting that some regulator other than mTOR signaling, for example, Akt, might also suppress the autophagic-lysosomal system. From these results, we suggested that Lys suppressed the activity of the autophagic-lysosomal system in part through activation of mTOR and reduced myofibrillar protein degradation in C2C12 myotubes.

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Animals
  • Autophagy / drug effects*
  • Carrier Proteins / metabolism
  • Cell Cycle Proteins
  • Eukaryotic Initiation Factors
  • Gene Expression Regulation / drug effects
  • Lysine / pharmacology*
  • Lysosomes / metabolism*
  • Methylhistidines / pharmacology
  • Mice
  • Microtubule-Associated Proteins / metabolism
  • Muscle Fibers, Skeletal / drug effects
  • Muscle Fibers, Skeletal / metabolism*
  • Muscle Proteins / genetics
  • Muscle Proteins / metabolism
  • Myofibrils / drug effects
  • Myofibrils / metabolism
  • Phosphoproteins / metabolism
  • Phosphorylation / drug effects
  • Proteasome Endopeptidase Complex / metabolism
  • Proteolysis / drug effects*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • TOR Serine-Threonine Kinases / metabolism
  • Time Factors
  • Tripartite Motif Proteins
  • Ubiquitin / metabolism
  • Ubiquitin-Protein Ligases / genetics
  • Ubiquitin-Protein Ligases / metabolism


  • Adaptor Proteins, Signal Transducing
  • Carrier Proteins
  • Cell Cycle Proteins
  • Eif4ebp1 protein, mouse
  • Eukaryotic Initiation Factors
  • Map1lc3b protein, mouse
  • Methylhistidines
  • Microtubule-Associated Proteins
  • Muscle Proteins
  • Phosphoproteins
  • RNA, Messenger
  • Tripartite Motif Proteins
  • Ubiquitin
  • Trim63 protein, mouse
  • Ubiquitin-Protein Ligases
  • TOR Serine-Threonine Kinases
  • Proteasome Endopeptidase Complex
  • Lysine