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Functional partnership between amphiphysin and dynamin in clathrin-mediated endocytosis

Nature Cell Biology volume 1, pages 33–39 (1999)Cite this article

Abstract

Amphiphysin, a protein that is highly concentrated in nerve terminals, has been proposed to function as a linker between the clathrin coat and dynamin in the endocytosis of synaptic vesicles. Here, using a cell-free system, we provide direct morphological evidence in support of this hypothesis. Unexpectedly, we also find that amphiphysin-1, like dynamin-1, can transform spherical liposomes into narrow tubules. Moreover, amphiphysin-1 assembles with dynamin-1 into ring-like structures around the tubules and enhances the liposome-fragmenting activity of dynamin-1 in the presence of GTP. These results show that amphiphysin binds lipid bilayers, indicate a potential function for amphiphysin in the changes in bilayer curvature that accompany vesicle budding, and imply a close functional partnership between amphiphysin and dynamin in endocytosis.

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Figure 1: Effect of amphiphysin-1 on the coupling between clathrin-coated budding and dynamin-mediated tubulation in protein-free liposomes composed of a brain lipid extract.
Figure 2: Amphiphysin alone tubulates liposomes composed of a brain lipid extract.
Figure 3: Amphiphysin-1 and dynamin-1 co-assemble into a hybrid coat.
Figure 4: Dynamin-1 and amphiphysin-1 co-polymerize into rings in a buffer of physiological ionic strength and pH.
Figure 5: GTP-dependent liposome fragmentation produced by dynamin-1 alone or by amphiphysin-1 and dynamin-1 together.
Figure 6: Effect of amphiphysin-1 on the recruitment of dynamin-1 to liposomes and on its GTPase activity.
Figure 7: Interactions of the amphiphysin dimer at a clathrin-coated endocytotic pit.

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Acknowledgements

This work was supported in part by grants from the NIH and the US Army Medical Research and Development Command (to P.D.C.), and a long-term fellowship from the Human Frontier Science Program (to V.H.).

Correspondence and requests for materials should be addressed to P.D.C.

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Author notes

  1. Kohji Takei and Vladimir I. Slepnev: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Cell Biology and Howard Hughes Medical Institute, Yale University School of Medicine, 295 Congress Avenue, New Haven, 06510, Connecticut , USA

    Kohji Takei, Vladimir I. Slepnev, Volker Haucke & Pietro De Camilli

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Correspondence to Pietro De Camilli.

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Takei, K., Slepnev, V., Haucke, V. et al. Functional partnership between amphiphysin and dynamin in clathrin-mediated endocytosis. Nat Cell Biol 1, 33–39 (1999). https://doi.org/10.1038/9004

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