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Nucleotide-dependent conformational changes in dynamin: evidence for a mechanochemical molecular spring

Nature Cell Biology volume 1, pages 27–32 (1999)Cite this article

Abstract

The GTPase dynamin plays an essential part in endocytosis by catalysing the fission of nascent clathrin-coated vesicles from the plasma membrane. Using preformed phosphatidylinositol-4,5-bisphosphate-containing lipid nanotubes as a membrane template for dynamin self-assembly, we investigate the conformational changes that arise during GTP hydrolysis by dynamin. Electron microscopy reveals that, in the GTP-bound state, dynamin rings appear to be tightly packed together. After GTP hydrolysis, the spacing between rings increases nearly twofold. When bound to the nanotubes, dynamin"s GTPase activity is cooperative and is increased by three orders of magnitude compared with the activity of unbound dynamin. An increase in the kcat (but not the Km) of GTP hydrolysis accounts for the pronounced cooperativity. These data indicate that a novel, lengthwise (‘spring-like’) conformational change in a dynamin helix may participate in vesicle fission.

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Figure 1: Purified dynamin associates with lipid nanotubes containing PtdIns(4,5)P2 and exhibits high GTPase activity.
Figure 2: The conformational states of dynamin in the presence of different nucleotides.
Figure 3: High rates of GTP hydrolysis by tubule-associated dynamin.
Figure 4: Cooperativity and V-type allostery of dynamin’s GTPase activity.
Figure 5: Potential involvement of stacks of rings in optimal GTP hydrolysis.
Figure 6: Model for a concerted helical expansion that drives endocytosis.

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Acknowledgements

We thank M. M. Kozlov for discussions and providing unpublished data, and N. Unwin for encouragement and discussions. This work was supported by the MRC and an NSF-NATO Postdoctoral Fellowship (M.H.B.S.).

Correspondence and requests for materials should be addressed to H.T.M.

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  1. Michael H. B. Stowell and Bruno Marks: These authors contributed equally to this work

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  1. MRC Laboratory of Molecular Biology, Hills Road, Cambridge , CB2 2QH, UK

    Michael H. B. Stowell, Bruno Marks, Patrick Wigge & Harvey T. McMahon

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Correspondence to Harvey T. McMahon.

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Stowell, M., Marks, B., Wigge, P. et al. Nucleotide-dependent conformational changes in dynamin: evidence for a mechanochemical molecular spring. Nat Cell Biol 1, 27–32 (1999). https://doi.org/10.1038/8997

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