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Measuring Rab GTPase-Activating Protein (GAP) Activity in Live Cells and Extracts

  • Protocol
Rab GTPases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1298))

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Abstract

Mammalian cells encode a diverse set of Rab GTPases and their corresponding regulators. In vitro biochemical screening has proven invaluable in assigning particular Rabs as substrates for their cognate GTPase-activating proteins. However, in vitro activity does not always reflect substrate specificity in cells. This method describes a functional test of GAP activity in cells or extracts that takes into account the presence of other factors or conditions that might change observed in vitro specificity.

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Acknowledgement

This research was supported by a grant from the National Institutes of Health (NIDDK 37332-28) to SRP.

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Authors and Affiliations

  1. Department of Biochemistry, Stanford University School of Medicine, 291 Campus Drive, Stanford, CA, 94305-5307, USA

    Ryan M. Nottingham & Suzanne R. Pfeffer

  2. Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX, 78712, USA

    Ryan M. Nottingham

Authors
  1. Ryan M. Nottingham
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  2. Suzanne R. Pfeffer
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Corresponding author

Correspondence to Suzanne R. Pfeffer .

Editor information

Editors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Peggy and Charles Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA

    Guangpu Li

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Nottingham, R.M., Pfeffer, S.R. (2015). Measuring Rab GTPase-Activating Protein (GAP) Activity in Live Cells and Extracts. In: Li, G. (eds) Rab GTPases. Methods in Molecular Biology, vol 1298. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2569-8_5

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  • DOI: https://doi.org/10.1007/978-1-4939-2569-8_5

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2568-1

  • Online ISBN: 978-1-4939-2569-8

  • eBook Packages: Springer Protocols

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