Studying the ArfGAP-Dependent Conformational Changes in SNAREs

  • Fernanda Rodriguez
  • Anne Spang
Part of the Methods in Molecular Biology book series (MIMB, volume 619)


Vesicle SNAREs (v-SNAREs) are included with high fidelity into each transport vesicle generated in the cell. These SNAREs determine the fate of vesicles, as they are the key factors deciding with which compartment a particular vesicle will fuse. The mechanism of high fidelity inclusion of SNAREs into transport vesicles is very difficult to study in vivo. Therefore, we use in vitro assays aiming to recapitulate SNARE uptake into vesicles. One of the key assays is a pull-down with SNARE tails fused to GST in the presence or absence of ArfGAPs and coat components such as the small GTPase Arf1 and coatomer. This in vitro assay allowed us to show that the ArfGAPs Glo3 and Gcs1 can induce a conformational change in SNAREs. Protease protection assays were used to confirm the conformational change and can also be used to address the question about the nature of the different conformations in SNARE proteins.

Key words

SNAREs Glo3p Arf1p conformational changes SNARE binding assay protease protection assays 



The work on ArfGAPs and SNAREs in the Spang lab is funded by the University of Basel and the Swiss National Science Foundation.


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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Fernanda Rodriguez
    • 1
  • Anne Spang
    • 1
  1. 1.Biozentrum, Growth and DevelopmentUniversity of BaselBaselSwitzerland

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