Puente Lawrence G, Voisin Sébastien, Lee Robin E C, Megeney Lynn A
Ottawa Health Research Institute, Molecular Medicine Program, Ottawa Hospital, and Department of Cellular and Molecular Medicine, Centre for Neuromuscular Disease, Faculty of Medicine, University of Ottawa, Ontario K1H 8L6, Canada.
Mol Cell Proteomics. 2006 Dec;5(12):2244-51. doi: 10.1074/mcp.M600134-MCP200. Epub 2006 Sep 13.
Multiple kinase activities are required for skeletal muscle differentiation. However, the mechanisms by which these kinase pathways converge to coordinate the myogenic process are unknown. Using multiple phosphoprotein and phosphopeptide enrichment techniques we obtained phosphopeptides from growing and differentiating C2C12 muscle cells and determined specific peptide sequences using LC-MS/MS. To place these phosphopeptides into a rational context, a bioinformatics approach was used. Phosphorylation sites were matched to known site-specific and to site non-specific kinase-substrate interactions, and then other substrates and upstream regulators of the implicated kinases were incorporated into a model network of protein-protein interactions. The model network implicated several kinases of known relevance to myogenesis including AKT, GSK3, CDK5, p38, DYRK, and MAPKAPK2 kinases. This combination of proteomics and bioinformatics technologies should offer great utility as the volume of protein-protein and kinase-substrate information continues to increase.
骨骼肌分化需要多种激酶活性。然而，这些激酶途径汇聚以协调肌生成过程的机制尚不清楚。我们使用多种磷酸化蛋白和磷酸肽富集技术，从生长和分化的C2C12肌肉细胞中获得磷酸肽，并使用液相色谱-串联质谱法（LC-MS/MS）确定特定的肽序列。为了将这些磷酸肽置于合理的背景中，我们采用了生物信息学方法。将磷酸化位点与已知的位点特异性和非特异性激酶-底物相互作用进行匹配，然后将所涉及激酶的其他底物和上游调节因子纳入蛋白质-蛋白质相互作用的模型网络中。该模型网络涉及几种已知与肌生成相关的激酶，包括AKT、GSK3、CDK5、p38、DYRK和MAPKAPK2激酶。随着蛋白质-蛋白质和激酶-底物信息数量的不断增加，这种蛋白质组学和生物信息学技术的结合应具有很大的实用性。
