Preliminary quantitative profile of differential protein expression between rat L6 myoblasts and myotubes by stable isotope labeling with amino acids in cell culture

Ziyou Cui, Xiulan Chen, Bingwen Lu, Kyu Park Sung, Tao Xu, Zhensheng Xie, Peng Xue, Junjie Hou, Haiying Hang, John R. Yates, Fuquan Yang

Research output: Contribution to journalArticlepeer-review

42 Scopus citations

Abstract

Defining the mechanisms governing myogenesis has advanced in recent years. Skeletal-muscle differentiation is a multi-step process controlled spatially and temporally by various factors at the transcription level. To explore those factors involved in myogenesis, stable isotope labeling with amino acids in cell culture (SILAC), coupled with high-accuracy mass spectrometry (LTQ-Orbitrap), was applied successfully. Rat L6 cell line is an excellent model system for studying muscle myogenesis in vitro. When mononucleate L6 myoblast cells reach confluence in culture plate, they could transform into multinucleate myotubes by serum starvation. By comparing protein expression of L6 myoblasts and terminally differentiated multinucleated myotubes, 1170 proteins were quantified and 379 proteins changed significantly in fully differentiated myotubes in contrast to myoblasts. These differentially expressed proteins are mainly involved in inter-or intracellular signaling, protein synthesis and degradation, protein folding, cell adhesion and extracelluar matrix, cell structure and motility, metabolism, substance transportation, etc. These findings were supported by many previous studies on myogenic differentiation, of which many up-regulated proteins were found to be involved in promoting skeletal muscle differentiation for the first time in our study. Insummary, our results provide new clues for understanding the mechanism of myogenesis.

Original languageEnglish (US)
Pages (from-to)1274-1292
Number of pages19
JournalProteomics
Volume9
Issue number5
DOIs
StatePublished - Mar 2009

Keywords

  • 2D-LC-LTQ-orbitrap
  • Quantitative proteomics
  • SILAC
  • Skeletal-muscle differentiation

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