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Proteomic characterization of the human centrosome by protein correlation profiling

Nature volume 426, pages 570–574 (2003)Cite this article

Abstract

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 cells1,2,3,4,5. 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.

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Figure 1: Proteomic analysis of the centrosome.
Figure 2: U2OS cells expressing a selection of novel centrosomal proteins as GFP fusion proteins. γ-Tubulin (centrosome marker, a–d) or α-tubulin (microtubule marker, e, f) was stained with Texas red and DNA with 4,6-diamidino-2-phenylindole; yellow indicates coincidence of green and red signals.
Figure 3: Protein correlation profiling of the human centrosome.

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Acknowledgements

We thank members of the Center for Experimental BioInformatics (CEBI) and the Nigg group for critical reading of the manuscript and fruitful discussions and M. Matzner for excellent technical support. We also thank M. J. Pebusque (INSERM U119, Marseille, France) for the kind gift of a cDNA clone of FOP. CEBI is supported by a grant by the Danish Natural Research Foundation. E.A.N. acknowledges support from the Max-Planck Society and the ‘Fonds der Chemischen Industrie’.

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Author notes

  1. Thibault Mayor

    Present address: Division of Biology MC 156-29, California Institute of Technology, 1200 East California Blvd, Pasadena, California, 91125, USA

  2. Jens S. Andersen and Christopher J. Wilkinson: These authors contributed equally to this work

Authors and Affiliations

  1. Center for Experimental BioInformatics (CEBI), Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, DK-5230 M, Odense, Denmark

    Jens S. Andersen, Peter Mortensen & Matthias Mann

  2. Department of Cell Biology, Max Planck Institute for Biochemistry, D-82152, Martinsried, Germany

    Christopher J. Wilkinson, Thibault Mayor & Erich A. Nigg

Authors
  1. Jens S. Andersen
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Correspondence to Erich A. Nigg or Matthias Mann.

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Andersen, J., Wilkinson, C., Mayor, T. et al. Proteomic characterization of the human centrosome by protein correlation profiling. Nature 426, 570–574 (2003). https://doi.org/10.1038/nature02166

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