Comparative proteomes of the proliferating C2C12 myoblasts and fully differentiated myotubes reveal the complexity of the skeletal muscle differentiation program
Comparative proteomes of the proliferating C2C12 myoblasts and fully differentiated myotubes reveal the complexity of the skeletal muscle differentiation program
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DOI：
10.1074/mcp.m400020-mcp200
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发表时间：
2004-11-01
影响因子：
7
通讯作者：
Raghow, R
中科院分区：
文献类型：
--
作者：
Tannu, NS;Rao, VK;Raghow, R
当在含低血清的生长培养基中培养时，小鼠C2 C12细胞退出细胞周期并经历明确的分化程序，最终形成肌球蛋白重链阳性的真正多核肌细胞。为了理解这一过程，我们通过二维PAGE、定量PDQuest图像分析和MS的组合方法比较了来自增殖的C2 C12细胞和完全分化的肌管的总蛋白、膜富集蛋白和核富集蛋白以及磷蛋白。从C2 C12成肌细胞和肌管中提取的000种蛋白质揭示，绝大多数丰富的蛋白质似乎被归为基本的、管家的和结构的功能，并且它们的稳态水平保持相对恒定。与此相反，75个蛋白质的高度调节过程中的表型转换的快速分裂C2 C12成肌细胞完全分化，多核，有丝分裂后的肌管。我们发现，差异积累的26磷蛋白也发生在C2 C12成肌细胞转化成肌管。我们通过MALDI-TOF-MS和LC-ESI-四极杆离子阱MS/MS鉴定了差异表达的蛋白质。我们证明了超过100种蛋白质，其中一些蛋白质首次与肌肉分化相关，它们调节细胞间和细胞内信号传导、细胞形状、增殖、凋亡和基因表达，影响骨骼肌分化的机制。
When cultured in low serum-containing growth medium, the mouse C2C12 cells exit cell cycle and undergo a well-defined program of differentiation that culminates in the formation of myosin heavy chain-positive bona fide multinucleated muscle cells. To gain an understanding into this process, we compared total, membrane- and nuclear-enriched proteins, and phospho-proteins from the proliferating C2C12 cells and the fully differentiated myotubes by the combined methods of two-dimensional PAGE, quantitative PDQuest image analysis, and MS. Quantification of more than 2,000 proteins from C2C12 myoblasts and myotubes revealed that a vast majority of the abundant proteins appear to be relegated to the essential, housekeeping and structural functions, and their steady state levels remain relatively constant. In contrast, 75 proteins were highly regulated during the phenotypic conversion of rapidly dividing C2C12 myoblasts into fully differentiated, multi-nucleated, post-mitotic myotubes. We found that differential accumulation of 26 phosphoproteins also occurred during conversion of C2C12 myoblasts into myotubes. We identified the differentially expressed proteins by MALDI-TOF-MS and LC-ESI-quadrupole ion trap MS/MS. We demonstrate that more than 100 proteins, some shown to be associated with muscle differentiation for the first time, that regulate inter- and intracellular signaling, cell shape, proliferation, apoptosis, and gene expression impinge on the mechanism of skeletal muscle differentiation.