Abstract
Protein coats deform flat lipid membranes into buds and capture membrane proteins to form transport vesicles1,2,3. The assembly/disassembly cycle of the COPI coat on Golgi membranes is coupled to the GTP/GDP cycle of the small G protein Arf1. At the heart of this coupling is the specific interaction of membrane-bound Arf1–GTP with coatomer, a complex of seven proteins that forms the building unit of the COPI coat4,5,6,7. Although COPI coat disassembly requires the catalysis of GTP hydrolysis in Arf1 by a specific GTPase-activating protein (ArfGAP1)8,9,10, the precise timing of this reaction during COPI vesicle formation is not known. Using time-resolved assays for COPI dynamics on liposomes of controlled size, we show that the rate of ArfGAP1-catalysed GTP hydrolysis in Arf1 and the rate of COPI disassembly increase over two orders of magnitude as the curvature of the lipid bilayer increases and approaches that of a typical transport vesicle. This leads to a model for COPI dynamics in which GTP hydrolysis in Arf1 is organized temporally and spatially according to the changes in lipid packing induced by the coat.
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Acknowledgements
We thank P. Randazzo and R. Premont for proteins and constructs, J. Moelleken for advice on coatomer purification and lipopeptide synthesis and M. Chabre for discussions. This work was supported by the CNRS (ACI ‘Jeune Chercheur’) and the EMBO Young Investigator Programme.
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Bigay, J., Gounon, P., Robineau, S. et al. Lipid packing sensed by ArfGAP1 couples COPI coat disassembly to membrane bilayer curvature. Nature 426, 563–566 (2003). https://doi.org/10.1038/nature02108
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DOI: https://doi.org/10.1038/nature02108
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