Proteomic characterization of the human centrosome by protein correlation profiling
Proteomic characterization of the human centrosome by protein correlation profiling
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DOI：
10.1038/nature02166
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发表时间：
2003-12-04
期刊：
影响因子：
64.8
通讯作者：
Mann, M
中科院分区：
文献类型：
--
作者：
Andersen, JS;Wilkinson, CJ;Mann, M
中心体是动物细胞的主要微管组织中心，通过其对细胞骨架的影响，参与细胞的形状、极性和运动。它在细胞分裂中也有重要的功能，因为它决定有丝分裂纺锤体的极点，在分裂的细胞之间分离复制的染色体（1-5）。尽管这种细胞器对细胞生物学的重要性和100多年的研究，其功能的许多方面仍然是谜，其结构和组成在很大程度上仍然未知。我们对细胞周期间期的人类中心体进行了基于质谱的蛋白质组学分析，通过对数百种蛋白质进行了定量分析。真正的中心体蛋白是通过与已知中心体蛋白的相关性和体内定位来揭示的。我们鉴定并验证了23种新成分，并在大量非特异性蛋白质的背景下鉴定了41种可能的候选成分以及绝大多数已知的中心体蛋白。蛋白质相关分析允许分析任何多蛋白复合物，可以通过分离富集，但不能纯化到均匀性。
The centrosome is the major microtubule-organizing centre of animal cells and through its influence on the cytoskeleton is involved in cell shape, polarity and motility. It also has a crucial function in cell division because it determines the poles of the mitotic spindle that segregates duplicated chromosomes between dividing cells(1-5). Despite the importance of this organelle to cell biology and more than 100 years of study, many aspects of its function remain enigmatic and its structure and composition are still largely unknown. We performed a mass-spectrometry-based proteomic analysis of human centrosomes in the interphase of the cell cycle by quantitatively profiling hundreds of proteins across several centrifugation fractions. True centrosomal proteins were revealed by both correlation with already known centrosomal proteins and in vivo localization. We identified and validated 23 novel components and identified 41 likely candidates as well as the vast majority of the known centrosomal proteins in a large background of nonspecific proteins. Protein correlation profiling permits the analysis of any multiprotein complex that can be enriched by fractionation but not purified to homogeneity.