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Research

Proteome Dynamics during C2C12 Myoblast Differentiation^*,S

Thomas Kislinger^,, Anthony O. Gramolini^,¶,||, Yan Pan^,¶,**, Khaled Rahman^,, David H. MacLennan^¶ and Andrew Emili^,¶,

From the Program in Proteomics and Bioinformatics, University of Toronto, Toronto, Ontario M5S 3E2, Canada and ^¶ the Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario M5G 1L6, Canada These authors contributed equally to this work ^|| Postdoctoral research fellow of the Heart and Stroke Foundation of Canada ^** Postdoctoral research fellow of the Heart and Stroke Foundation of Ontario. Present address: Protana Inc., 251 Attwell Dr., Toronto, Ontario M9W TH4, Canada Present address: Dept. of Biochemistry and Molecular Biology, University of Calgary, 3330 Hospital Dr., Calgary, Alberta T2N 4N1, Canada

Mouse-derived C2C12 myoblasts serve as an experimentally tractable model system for investigating the molecular basis of skeletal muscle cell specification and development. To examine the biochemical adaptations associated with myocyte formation comprehensively, we used large scale gel-free tandem mass spectrometry to monitor global proteome alterations throughout a time course analysis of the myogenic C2C12 differentiation program. The relative abundance of 1,800 high confidence proteins was tracked across multiple time points using capillary scale multidimensional liquid chromatography coupled to high throughput shotgun sequencing. Hierarchical clustering of the resulting profiles revealed differential waves of expression of proteins linked to intracellular signaling, transcription, cytoarchitecture, adhesion, metabolism, and muscle contraction across the early, mid, and late stages of differentiation. Several hundred previously uncharacterized proteins were likewise detected in a stage-specific manner, suggesting novel roles in myogenesis and/or muscle function. These proteomic data are complementary to recent microarray-based studies of gene expression patterns in developing myotubes and provide a holistic framework for understanding how diverse biochemical processes are coordinated at the cellular level during skeletal muscle development.

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