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Rho GTPases pp 107-119 | Cite as

A Quantitative Fluorometric Approach for Measuring the Interaction of RhoGDI with Membranes and Rho GTPases

  • Jared Johnson
  • Richard A. Cerione
  • Jon W. Erickson
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 827)

Abstract

Tight regulation of Rho GTPase-signaling functions requires the proper localization of proteins to the membrane and cytosolic compartments, which can themselves undergo reconfiguration in response to signaling events. The importance of lipid-mediated membrane signal transduction continues to emerge as a critical event in many Rho GTPase-signaling pathways. Here we describe methods for the reconstitution of lipid-modified Rho GTPases with defined lipid vesicles and how this system can be used as a real-time assay for monitoring protein–membrane interactions.

Key words

Cdc42 Rho Fluorescence resonance energy transfer Synthetic lipid vesicles Guanine nucleotide dissociation inhibitor 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Jared Johnson
    • 1
  • Richard A. Cerione
    • 1
    • 2
  • Jon W. Erickson
    • 1
  1. 1.Department of Chemistry and Chemical BiologyCornell UniversityIthacaUSA
  2. 2.Department of Molecular MedicineCornell UniversityIthacaUSA

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