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

Academic Article
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Overview

authors

  • Cui, Z. Y.
  • Chen, X. L.
  • Lu, B. W.
  • Park, S. K.
  • Xu, T.
  • Xie, Z. S.
  • Xue, P.
  • Hou, J. J.
  • Hang, H. Y.
  • Yates III, John
  • Yang, F. Q.

publication date

  • March 2009

journal

  • Proteomics  Journal

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 extracellular 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. In summary, our results provide new clues for understanding the mechanism of myogenesis.

subject areas

  • Amino Acids
  • Animals
  • Cell Line
  • Cell Physiological Processes
  • Chromatography, Liquid
  • Gene Expression Regulation
  • Isotope Labeling
  • Muscle Development
  • Muscle Fibers, Skeletal
  • Muscle Proteins
  • Myoblasts
  • Proteomics
  • Rats
  • Signal Transduction
  • Tandem Mass Spectrometry
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Research

keywords

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

PubMed Central ID

  • PMC2946197

International Standard Serial Number (ISSN)

  • 1615-9853

Digital Object Identifier (DOI)

  • 10.1002/pmic.200800354

PubMed ID

  • 19253283
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Additional Document Info

start page

  • 1274

end page

  • 1292

volume

  • 9

issue

  • 5

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