JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2014, Vol. 52 ›› Issue (12): 54-59.doi: 10.6040/j.issn.1671-7554.0.2014.146

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Centrosome isolation and analysis by mass spectrometry-based proteomics

ZHONG Kaize1,2,3, ZHAO Jian2, YUAN Shoudao4, WANG Luyu3, JING Yanjie3, HAO Tong3, CHEN Jianxing4, ZHAN Qimin5, JIANG Wei3,5, ZHU Changjun3, PAN Lina3   

  1. 1. Department of Thoracic Surgery, Tianjin First Center Hospital, Tianjin 300192, China;
    2. Department of Thoracic Surgery, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China;
    3. Key Laboratory of Molecular Cellular Systems Biology, College of Life Sciences, Tianjin Normal University, Tianjin 300387, China;
    4. The Cancer Research Center of Shandong University, Jinan 250012, Shandong, China;
    5. State Key Laboratory of Molecular Oncology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100021, China
  • Received:2014-03-17 Revised:2014-10-29 Published:2014-12-10

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Abstract: Objective To identify the composition of centrosome proteins by human centrosome extraction, purification and protein spectrum analysis. Methods Centrosomes in K562 cells were extracted through density gradient centrifugation. Protein spectrum analysis was carried out on the sample tested by Western blotting. System analysis of all these centrosome proteins were proceeded by Gene Ontology (GO). Results The human centrosome was successfully extracted and purified. Cell lysis fractions where centrosome-associated protein γ-Tubulin accumulated were detected. There were no mitochondrial protein VDAC1 and ribosomal protein RPL15 in these fractions. The centrosome proteincomponents, including cytoplasm material transport protein, cytoskeleton protein, membrane combining protein, kinase protein and many other protein molecules, covered a variety of the biological message pathways within cells. Conclusion Centrosomes in different cell growth periods can be extracted and purified through this method. Combining protein spectrum analysis, this study is helpful to understand the basic structure and functions of the centosome.

Key words: Mass spectrometry, Centrosome, Proteomics

CLC Number: 

  • Q291
[1] Doxsey S, Zimmerman W, Mikule K. Centrosome control of the cell cycle[J]. Trends Cell Biol, 2005, 15(6): 303-311.
[2] Labbé J C, McCarthy E K, Goldstein B. The forces that position a mitotic spindle asymmetrically are tethered until after the time of spindle assembly[J]. J Cell Biol, 2004, 167(2): 245-256.
[3] Nigg E A, Stearns T. The centrosome cycle: centriole biogenesis, duplication and inherent asymmetries[J]. Nat Cell Biol, 2011, 13(10): 1154-1160.
[4] Pihan G A. Centrosome dysfunction contributes to chromosome instability, chromoanagenesis, and genome reprograming in cancer[J]. Front Oncol, 2013, 12(3):277.
[5] Nigg E A, Raff J W. Centrioles, centrosomes and cilia in health and disease[J]. Cell, 2009,139(4): 663-678.
[6] Nogales-Cadenas R, Abascal F, Díez-Pérez J, et al. CentrosomeDB: a human centrosomal proteins database[J]. Nucleic Acids Res, 2009, 37: D175-D180.
[7] Tomasello M F, Guarino F, Reina S, et al.The voltage-dependent anion selective channel 1 (VDAC1) topography in the mitochondrial outer membrane as detected in intact cell[J]. PLoS One, 2013, 8(12): 81522.
[8] Simoff I, Moradi H, Nygård O. Functional characterization of ribosomal protein L15 from Saccharomyces cerevisiae[J].Curr Genet, 2009, 55(2): 111-125.
[9] Consortium G O. The gene ontology project in 2008[J]. Nucleic Acids Res, 2008, 36: D440-D444.
[10] Jakobsen L, Vanselow K, Skogs M, et al. Novel asymmetrically localizing components of human centrosomes identified by complementary proteomics methods[J]. EMBO J, 2011, 30(8):1520-1535.
[11] Pihan G A, Wallace J, Zhou Y, et al. Centrosome abnormalities and chromosome instability occur together in pre-invasive carcinomas[J]. Cancer Res, 2003, 63(6): 1398-1404.
[12] Andersen J S, Wilkinson C J, Mayor T, et al. Proteomic characterization of the human centrosome by protein correlation profiling[J]. Nature, 2003, 426(6966): 570-574.
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