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Liver Proteomics pp 125–140Cite as

Quantitative Analysis of Liver Golgi Proteome in the Cell Cycle

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 909))

Abstract

During mitosis, the Golgi membranes in mammalian cells undergo a continuous disassembly process and generate mitotic fragments that are distributed into the daughter cells and reassembled into new Golgi after mitosis. This disassembly and reassembly process is critical for Golgi biogenesis during cell division, but the underlying molecular mechanism is poorly understood. In this study, we have recapitulated this process using an in vitro assay and analyzed the proteins that are associated with interphase and mitotic Golgi membranes using quantitative proteomics that combines the isobaric tags for relative and absolute quantification approach with OFFGEL isoelectric focusing separation and LC-MALDI-MS/MS. A total of 1,193 Golgi-associated proteins were identified and quantified. These included broad functional categories: Golgi structural proteins, Golgi resident enzymes, SNAREs, Rab GTPases, and secretory and cytoskeletal proteins. More importantly, the combination of the quantitative proteomic approach with Western blot analysis allowed us to unveil 86 proteins with significant changes in abundance under the mitotic condition compared to the interphase condition. Altogether, this systematic quantitative proteomic study revealed candidate proteins of the molecular machinery that controls the Golgi disassembly and reassembly processes in the cell cycle.

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Acknowledgements

We gratefully acknowledge Drs. F. Gorelick, K. Gull, T. Kreis, M. Lowe, K. Moremen, A. Price, A. Satoh, J. Seemann, D. Sheff, D. Shields, and G. Warren for generously providing antibodies. We thank J. Williams and D. Tang for suggestions and reagents. We thank Sarah Volk for her assistance in OFFGEL electrophoresis. This work was supported by National Institute of Health grant P41 RR018627 to P. Andrews, and was partially supported by the National Institutes of Health (GM087364) and the American Cancer Society (RGS-09-278-01-CSM) to Y.W.

Author information

Authors and Affiliations

  1. Department of Physiology, Wayne State University, Detroit, MI, USA

    Xuequn Chen

  2. Department of Biological Chemistry, The University of Michigan, Ann Arbor, MI, USA

    Philip C. Andrews

  3. Department of Molecular, Cellular and Developmental Biology, The University of Michigan, Ann Arbor, MI, USA

    Yanzhuang Wang

Authors
  1. Xuequn Chen
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  2. Philip C. Andrews
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  3. Yanzhuang Wang
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Corresponding author

Correspondence to Yanzhuang Wang .

Editor information

Editors and Affiliations

  1. Warren Alpert Medical School, Proteomics Core Facility, Brown University, Hoppin Street 1, Providence, 02903, Rhode Island, USA

    Djuro Josic

  2. Div. Hematology & Oncology, Rhode Island Hospital, Eddy St. 593, Providence, 02903, Rhode Island, USA

    Douglas C. Hixson

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© 2012 Springer Science+Business Media New York

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Chen, X., Andrews, P.C., Wang, Y. (2012). Quantitative Analysis of Liver Golgi Proteome in the Cell Cycle. In: Josic, D., Hixson, D. (eds) Liver Proteomics. Methods in Molecular Biology, vol 909. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-959-4_9

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  • DOI: https://doi.org/10.1007/978-1-61779-959-4_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-958-7

  • Online ISBN: 978-1-61779-959-4

  • eBook Packages: Springer Protocols

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