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Proteomic and Biochemical Methods to Study the Cytoskeletome

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Adhesion Protein Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1046))

Abstract

The cytoskeleton is fundamental to many cellular functions including cell proliferation, differentiation, adhesion, and migration. It is composed of actin, microtubules, intermediate filaments, and integrin cell surface receptors, which form focal adhesions with the extracellular matrix. These elements are highly integrated in the cell providing a rigid network of interconnected cables and protein scaffolds, which generate force and mechanical support to maintain cell shape and movement. However, the cytoskeleton is not just a simple compilation of static filaments that dictate cell adhesion and morphology—it is highly plastic with the inherent ability to assemble and disassemble in response to diverse and complex cellular cues. Thus, biochemical and proteomic methods are needed to better understand the cytoskeleton network and its dynamic signal transduction functions in health and disease. This chapter describes methods for the biochemical enrichment and mass spectrometry-based proteomic analyses of the cytoskeletome. We also detail how these methods can be used to investigate the cytoskeletome of migrating cells and their purified pseudopodia membrane projections.

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Acknowledgments

This work was supported by the NIH-IRACDA (NIH—Institutional Research and Academic Career Development Award) Postdoctoral Fellowship GM06852 (J.A. Kelber) and NIH grants CA097022 and CA129231 (R.L. Klemke).

Author information

Authors and Affiliations

  1. Department of Pathology and Moores Cancer Center, University of California, San Diego, La Jolla, CA, USA

    Richard L. Klemke, Xinning Jiang & Sunkyu Choi

  2. Department of Biology, California State University, Northridge, Northridge, CA, USA

    Jonathan A. Kelber

Authors
  1. Richard L. Klemke
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  2. Xinning Jiang
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  3. Sunkyu Choi
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  4. Jonathan A. Kelber
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Editor information

Editors and Affiliations

  1. , Department of Oncology, University of Oxford, Roosevelt Drive, Oxford, OX3 7DQ, United Kingdom

    Amanda S. Coutts

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Klemke, R.L., Jiang, X., Choi, S., Kelber, J.A. (2013). Proteomic and Biochemical Methods to Study the Cytoskeletome. In: Coutts, A. (eds) Adhesion Protein Protocols. Methods in Molecular Biology, vol 1046. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-538-5_12

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  • DOI: https://doi.org/10.1007/978-1-62703-538-5_12

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-537-8

  • Online ISBN: 978-1-62703-538-5

  • eBook Packages: Springer Protocols

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