Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
Proteomics. 2010 Nov;10(22):3935-56. doi: 10.1002/pmic.201000289. Epub 2010 Nov 2.
Developments in subcellular fractionation strategies have provided the means to profile and analyze the protein composition of organelles and cellular structures by proteomics. Here, we review the application of classical (e.g. density gradient centrifugation) and emerging sophisticated techniques (fluorescent-assisted organelle sorting) in the fractionation, and statistical/bioinformatics tools for the prediction of protein localization in subcellular proteomics. We also review the validation methods currently used (such as microscopy, RNA interference and multiple reaction monitoring) and discuss the importance of verification of the results obtained in subcellular proteomics. Finally, the numerous challenges facing subcellular proteomics including the dynamics of organelles are being examined. However, complementary approaches such as modern statistics, bioinformatics and large-scale integrative analysis are beginning to emerge as powerful tools to proteomics for analyzing subcellular organelles and structures.
亚细胞分级分离策略的发展为蛋白质组学提供了通过分析亚细胞蛋白质组成来描绘和分析细胞器和细胞结构的方法。在这里，我们回顾了经典（例如密度梯度离心）和新兴复杂技术（荧光辅助细胞器分选）在分级分离中的应用，以及用于预测亚细胞蛋白质组学中蛋白质定位的统计/生物信息学工具。我们还回顾了目前使用的验证方法（例如显微镜，RNA 干扰和多重反应监测），并讨论了验证亚细胞蛋白质组学获得的结果的重要性。最后，正在检查亚细胞蛋白质组学面临的许多挑战，包括细胞器的动态。但是，互补方法，如现代统计学，生物信息学和大规模综合分析，正开始成为分析亚细胞细胞器和结构的蛋白质组学的强大工具。
