期刊
EMBO JOURNAL
卷 30, 期 8, 页码 1520-1535出版社
WILEY
DOI: 10.1038/emboj.2011.63
关键词
centrosome; mass spectrometry-based proteomics; mother and daughter centriole; protein turnover; SILAC
资金
- Lundbeck Foundation
- Danish Agency for Science, Technology and Innovation
- European Commission [HEALTH-F4-2008-201648/PROSPECTS]
- Max Planck Society
- BMBF [01GS0859]
- Knut and Alice Wallenberg foundation
- HPA project
Centrosomes in animal cells are dynamic organelles with a proteinaceous matrix of pericentriolar material assembled around a pair of centrioles. They organize the microtubule cytoskeleton and the mitotic spindle apparatus. Mature centrioles are essential for biogenesis of primary cilia that mediate key signalling events. Despite recent advances, the molecular basis for the plethora of processes coordinated by centrosomes is not fully understood. We have combined protein identification and localization, using PCP-SILAC mass spectrometry, BAC transgeneOmics, and antibodies to define the constituents of human centrosomes. From a background of non-specific proteins, we distinguished 126 known and 40 candidate centrosomal proteins, of which 22 were confirmed as novel components. An antibody screen covering 4000 genes revealed an additional 113 candidates. We illustrate the power of our methods by identifying a novel set of five proteins preferentially associated with mother or daughter centrioles, comprising genes implicated in cell polarity. Pulsed labelling demonstrates a remarkable variation in the stability of centrosomal protein complexes. These spatiotemporal proteomics data provide leads to the further functional characterization of centrosomal proteins. The EMBO Journal (2011) 30, 1520-1535. doi:10.1038/emboj.2011.63; Published online 11 March 2011 Subject Categories: cell & tissue architecture; cell cycle; genomic & computational biology
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