Apr 22, 2020
Rab29 fast exchange mutants: characterization of a challenging Rab GTPase
- Rachel C. Gomez1,
- Edmundo G. Vides1,
- Suzanne R Pfeffer1
- 1Stanford University School of Medicine
External link: https://doi.org/10.7554/eLife.79771
Protocol Citation: Rachel C. Gomez, Edmundo G. Vides, Suzanne R Pfeffer 2020. Rab29 fast exchange mutants: characterization of a challenging Rab GTPase. protocols.io https://dx.doi.org/10.17504/protocols.io.bffrjjm6
Manuscript citation:
Rab GTPases: Methods and Protocols, Second Edition, Guangpu Li, Editor; Springer
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 22, 2020
Last Modified: April 22, 2020
Protocol Integer ID: 36049
Keywords: Nucleotide binding, Rab GTPase, fast exchange mutant, Rab29,
Abstract
Rab29 has been implicated in multiple membrane trafficking processes with no described effectors or regulating proteins. Its fast nucleotide exchange rate and inability to bind GDI in cytosol make it a unique and poorly understood Rab. Because the conventional, “GTP-locked” Rab mutation does not have the desired effect in Rab29, we present here the use of a fluorescence-based assay to characterize novel mutants that display faster nucleotide exchange rates, allowing for GEF-independent Rab29 activation.
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