Novel asymmetrically localizing components of human centrosomes identified by complementary proteomics methods
2011 (English)In: EMBO Journal, ISSN 0261-4189, E-ISSN 1460-2075, Vol. 30, no 8, 1520-1535 p.Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
2011. Vol. 30, no 8, 1520-1535 p.
centrosome, mass spectrometry-based proteomics, mother and daughter centriole, protein turnover, SILAC
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Cell and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-33978DOI: 10.1038/emboj.2011.63ISI: 000290306300013ScopusID: 2-s2.0-79955009072OAI: oai:DiVA.org:kth-33978DiVA: diva2:419919
FunderScience for Life Laboratory - a national resource center for high-throughput molecular bioscience
QC 201105302011-05-302011-05-232011-12-06Bibliographically approved