School of Chemistry and Chemical Engineering, Harbin Institute of Technology, Harbin 150090, Heilongjiang, PR China.
Department of Ophthalmology, The Second Hospital, Jilin University, Changchun 130012, Jilin, PR China.
Int J Biol Macromol. 2019 Mar 1;124:681-688. doi: 10.1016/j.ijbiomac.2018.11.265. Epub 2018 Nov 28.
Milk fat globule-EGF factor 8 (MFG-E8) is one of the major proteins in milk fat globule membrane. In this study, mouse-derived CC myoblast cells were served 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 myocytes formation comprehensively, a liquid chromatography coupled with tandem mass spectrometry label-free semi-quantitative approach was used to analyse the myogenic CC proliferation program. Over 1987 proteins were identified in CC cells. The MFG-E8 (200 μg/mL) and MFG-E8 (500 μg/mL) with significant differences were compared based on the relative abundance. The result profiles of regulation of MFG-E8 to the expression of proteins in CC cells revealed that differential waves of expression of proteins linked to intracellular signaling, transcription, cytoarchitecture, adhesion, metabolism, and muscle contraction across during the CC cell proliferation process. Based on the analysis of KEGG and STRING database, further to verification the expression of PI3K and ERK phosphorylation levels by Western blot. This study found that the data of proteomic was complementary to recent MFG-E8 studies of protein expression patterns in developing myotubes and provided a holistic framework for understanding how diverse biochemical processes are coordinated at the cellular level during skeletal muscle development.
乳脂肪球 EGF 因子 8(MFG-E8)是乳脂肪球膜中的主要蛋白质之一。在这项研究中,鼠源 CC 成肌细胞被用作研究骨骼肌细胞特化和发育的分子基础的实验上可处理的模型系统。为了全面研究与肌细胞形成相关的生化适应,采用液相色谱串联质谱非标记半定量方法分析了成肌 CC 增殖程序。在 CC 细胞中鉴定出超过 1987 种蛋白质。基于相对丰度,比较了 MFG-E8(200μg/mL)和 MFG-E8(500μg/mL)之间具有显著差异的蛋白质。MFG-E8 对 CC 细胞中蛋白质表达的调控的结果图谱表明,在 CC 细胞增殖过程中,与细胞内信号转导、转录、细胞结构、黏附、代谢和肌肉收缩相关的蛋白质表达存在差异波。通过对 KEGG 和 STRING 数据库的分析,并通过 Western blot 进一步验证 PI3K 和 ERK 磷酸化水平的表达。这项研究发现,蛋白质组学数据补充了最近关于发育中的肌管中 MFG-E8 蛋白质表达模式的研究,并为理解在骨骼肌发育过程中细胞水平上如何协调不同的生化过程提供了整体框架。