Gao Yan, Lim Teck Kwang, Lin Qingsong, Li Sam Fong Yau
Department of Chemistry, Faculty of Science, National University of Singapore, Singapore.
Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore.
Electrophoresis. 2016 May;37(10):1270-6. doi: 10.1002/elps.201500527. Epub 2016 Apr 4.
Many protein extraction methods have been developed for plant proteome analysis but information is limited on the optimal protein extraction method from algae species. This study evaluated four protein extraction methods, i.e. direct lysis buffer method, TCA-acetone method, phenol method, and phenol/TCA-acetone method, using green algae Chlorella vulgaris for proteome analysis. The data presented showed that phenol/TCA-acetone method was superior to the other three tested methods with regards to shotgun proteomics. Proteins identified using shotgun proteomics were validated using sequential window acquisition of all theoretical fragment-ion spectra (SWATH) technique. Additionally, SWATH provides protein quantitation information from different methods and protein abundance using different protein extraction methods was evaluated. These results highlight the importance of green algae protein extraction method for subsequent MS analysis and identification.
许多蛋白质提取方法已被开发用于植物蛋白质组分析，但关于从藻类物种中提取最佳蛋白质的方法的信息有限。本研究使用绿藻小球藻评估了四种蛋白质提取方法，即直接裂解缓冲液法、三氯乙酸-丙酮法、酚法和酚/三氯乙酸-丙酮法，用于蛋白质组分析。所呈现的数据表明，在鸟枪法蛋白质组学方面，酚/三氯乙酸-丙酮法优于其他三种测试方法。使用鸟枪法蛋白质组学鉴定的蛋白质通过所有理论碎片离子光谱的顺序窗口采集（SWATH）技术进行验证。此外，SWATH提供了来自不同方法的蛋白质定量信息，并评估了使用不同蛋白质提取方法的蛋白质丰度。这些结果突出了绿藻蛋白质提取方法对后续质谱分析和鉴定的重要性。
