Cheng Huamin, Zhao Hongwei, Yang Tinghan, Ruan Sunlan, Wang Haihua, Xiang Nan, Zhou Hailong, Li Qing X, Diao Xiaoping
State Key Laboratory of South China Sea Marine Resource Utilization, Hainan University, Haikou, P. R. China.
Institute of Tropical Agriculture and Forestry, Hainan University, Haikou, P. R. China.
Electrophoresis. 2018 Apr;39(8):1062-1070. doi: 10.1002/elps.201700436. Epub 2018 Feb 12.
Corals especially the reef-building species are very important to marine ecosystems. Proteomics has been used for researches on coral diseases, bleaching and responses to the environment change. A robust and versatile protein extraction protocol is required for coral proteomics. However, a comparative evaluation of different protein extraction protocols is still not available for proteomic analysis of stony corals. In the present study, five protocols were compared for protein extraction from stony corals. The five protocols were TRIzol, phenol-based extraction (PBE), trichloroacetic acid (TCA)-acetone, glass bead-assisted extraction (GBAE) and a commercially available kit. PBE, TRIzol and the commercial kit were more robust for extracting proteins from stony corals. The protein extraction efficiency and repeatability, two dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) were employed to evaluate the protocols. The results indicated that PBE protocol had the better protein extraction efficiency than the others. Protein extraction coverage varied among the procedures. Each protocol favored for certain proteins. Therefore, it is very important for coral proteomic analysis to select a suitable protein protocol upon the experimental design. In general, PBE protocol can be the first choice for extracting proteins from stony corals.
珊瑚，尤其是造礁珊瑚物种，对海洋生态系统非常重要。蛋白质组学已被用于珊瑚疾病、白化以及对环境变化响应的研究。珊瑚蛋白质组学需要一种强大且通用的蛋白质提取方案。然而，对于石珊瑚的蛋白质组学分析，目前尚无不同蛋白质提取方案的比较评估。在本研究中，比较了从石珊瑚中提取蛋白质的五种方案。这五种方案分别是TRIzol法、基于苯酚的提取法（PBE）、三氯乙酸（TCA）-丙酮法、玻璃珠辅助提取法（GBAE）和一种市售试剂盒。PBE法、TRIzol法和市售试剂盒在从石珊瑚中提取蛋白质方面表现更强健。采用蛋白质提取效率和重复性、二维电泳（2-DE）以及基质辅助激光解吸/电离飞行时间质谱（MALDI TOF MS）来评估这些方案。结果表明，PBE方案的蛋白质提取效率优于其他方案。不同提取程序的蛋白质提取覆盖率有所不同。每种方案都有利于某些特定蛋白质的提取。因此，对于珊瑚蛋白质组学分析而言，根据实验设计选择合适的蛋白质提取方案非常重要。总体而言，PBE方案可作为从石珊瑚中提取蛋白质的首选方案。
