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Rab GTPases pp 61-71 | Cite as

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

  • Ryan M. Nottingham
  • Suzanne R. Pfeffer
Part of the Methods in Molecular Biology book series (MIMB, volume 1298)

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.

Key words

GTPase-activating protein Rab GTPase Effector protein 

Notes

Acknowledgement

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

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of BiochemistryStanford University School of MedicineStanfordUSA
  2. 2.Department of Molecular Biosciences, Institute for Cellular and Molecular BiologyThe University of Texas at AustinAustinUSA

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