Proteome analysis of differentiating human myoblasts by dialysis-assisted two-dimensional gel electrophoresis (DAGE)
Proteome analysis of differentiating human myoblasts by dialysis-assisted two-dimensional gel electrophoresis (DAGE)
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DOI：
10.1002/pmic.200700261
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发表时间：
2008-01-01
期刊：
影响因子：
3.4
通讯作者：
Svinartchouk, Fedor
中科院分区：
文献类型：
--
作者：
Gonnet, Florence;Bouazza, Belaid;Svinartchouk, Fedor
在本研究中，通过透析辅助2-DE （page,[1]）研究了人成肌细胞分化过程中细胞质表达蛋白的修饰。分化第5天和第13天分析了约1000个斑点，蛋白表达动态范围超过1000倍。在成肌分化过程中，不匹配点的数量和匹配点之间的数量差异程度显著增加。通过MALDI-TOF ms鉴定了100多个差异表达点，并通过Western blot分析验证了8个蛋白的分化相关表达模式。几种蛋白的差异表达首次在人肌管中得到证实。有趣的是，匠心通路分析将这些蛋白质中的30个分成两个重叠的网络，其中包含IGF-1和肿瘤坏死因子作为主要节点，这两种蛋白质已知在细胞发生中起关键作用。我们的研究结果说明了人类成肌细胞分化过程中蛋白质组的大量重排，并为参与这一复杂过程的新伙伴提供了证据。
In the present study, modifications in cytosolic expressed proteins during human myoblast differentiation were studied by dialysis-assisted 2-DE (DAGE, [1]). About 1000 spots were analysed on the 5th and 13th day of differentiation with a dynamic range of protein expression exceeding 1000-fold. During myogenic differentiation, the number of nonmatching spots as well as the extent of quantitative differences between matched spots significantly increased. Over one hundred differentially expressed spots were excised and identified by MALDI-TOF MS. The differentiation-associated expression pattern of eight proteins was validated by Western blot analysis. Differential expression of several proteins was demonstrated for the first time in human myotubes. Interestingly, Ingenuity pathway analysis grouped 30 of these proteins into two overlapping networks containing as principal nodes IGF-1 and tumour necrosis factor, two proteins known to play a crucial role in cytogenesis. Our results illustrate the large rearrangement of the proteome during the differentiation of human myoblasts and provide evidence for new partners involved in this complex process.