Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021:2293:19-25.
doi: 10.1007/978-1-0716-1346-7_2.

Rab29 Fast Exchange Mutants: Characterization of a Challenging Rab GTPase

Rachel C Gomez  1 , Edmundo G Vides  1 , Suzanne R Pfeffer  2
Affiliations

Rab29 Fast Exchange Mutants: Characterization of a Challenging Rab GTPase

Rachel C Gomez et al. Methods Mol Biol. 2021.

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 Rab29 mutants (I64T and V156G) that display faster nucleotide exchange rates, allowing for GEF-independent Rab29 activation.

Keywords: Fast exchange mutant; MANT-GDP; Nucleotide exchange; Rab GTPase; Rab29.

PubMed Disclaimer

References

    1. Shoji I, Kikuchi A, Kuroda S, Takai Y (1989) Kinetic analysis of the binding of guanine nucleotide to bovine brain smg p25A. Biochem Biophys Res Commun 162(1):273–281. https://doi.org/10.1016/0006-291x(89)91992-x - DOI - PubMed
    1. Rak A, Pylypenko O, Durek T, Watzke A, Kushnir S, Brunsveld L, Waldmann H, Goody RS, Alexandrov K (2003) Structure of Rab GDP-dissociation inhibitor in complex with prenylated YPT1 GTPase. Science 302(5645):646–650. https://doi.org/10.1126/science.1087761 - DOI - PubMed
    1. Barr F, Lambright DG (2010) Rab GEFs and GAPs. Curr Opin Cell Biol 22(4):461–470. https://doi.org/10.1016/j.ceb.2010.04.007 - DOI - PubMed - PMC
    1. Bos JL, Rehmann H, Wittinghofer A (2007) GEFs and GAPs: critical elements in the control of small G proteins. Cell 129(5):865–877. https://doi.org/10.1016/j.cell.2007.05.018 - DOI - PubMed
    1. Gavriljuk K, Gazdag EM, Itzen A, Kötting C, Goody RS, Gerwert K (2012) Catalytic mechanism of a mammalian Rab·RabGAP complex in atomic detail. Proc Natl Acad Sci U S A 109(52):21348–21353. https://doi.org/10.1073/pnas.1214431110 - DOI - PubMed - PMC

LinkOut - more resources