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Membrane Trafficking pp 81–90Cite as

Investigating Signaling Processes in Membrane Trafficking

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1270))

Abstract

Signaling and phosphorylation can be very difficult areas to explore, as there can be a lot of cross-talk between signaling pathways, and the stoichiometry of phosphorylation is often very low, and is typically transient. Here we describe an innovative assay using an immunoprecipitation approach, followed by a kinase assay, coupled with a phosphorylated substrate-specific antibody. We also indicate a database and prediction program that can be used in these situations.

We apply these methods to investigate the regulation of ER-to-Golgi trafficking by protein phosphorylation of critical components in the trafficking machinery. Key components of this transport step are well known thanks to the pioneering work of the 2013 Nobel Prize winners James Rothman, Randy Schekman, and Thomas Südhof. However, the regulation aspect of this process is relatively unexplored.

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Authors and Affiliations

  1. School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, NSW, 2052, Australia

    Laura J. Sharpe Ph.D. & Andrew J. Brown Ph.D.

Authors
  1. Laura J. Sharpe Ph.D.
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  2. Andrew J. Brown Ph.D.
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Corresponding author

Correspondence to Andrew J. Brown Ph.D. .

Editor information

Editors and Affiliations

  1. Department of Biochemistry, National University of Singapore, Singapore, Singapore

    Bor Luen Tang

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Sharpe, L.J., Brown, A.J. (2015). Investigating Signaling Processes in Membrane Trafficking. In: Tang, B. (eds) Membrane Trafficking. Methods in Molecular Biology, vol 1270. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2309-0_6

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  • DOI: https://doi.org/10.1007/978-1-4939-2309-0_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2308-3

  • Online ISBN: 978-1-4939-2309-0

  • eBook Packages: Springer Protocols

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