Architectural and mechanistic insights into an EHD ATPase involved in membrane remodelling


The ability to actively remodel membranes in response to nucleotide hydrolysis has largely been attributed to GTPases of the dynamin superfamily, and these have been extensively studied1. Eps15 homology (EH)-domain-containing proteins (EHDs/RME-1/pincher) comprise a less-well-characterized class of highly conserved eukaryotic ATPases implicated in clathrin-independent endocytosis2, and recycling from endosomes3,4. Here we show that EHDs share many common features with the dynamin superfamily, such as a low affinity for nucleotides, the ability to tubulate liposomes in vitro, oligomerization around lipid tubules in ring-like structures and stimulated nucleotide hydrolysis in response to lipid binding. We present the structure of EHD2, bound to a non-hydrolysable ATP analogue, and provide evidence consistent with a role for EHDs in nucleotide-dependent membrane remodelling in vivo. The nucleotide-binding domain is involved in dimerization, which creates a highly curved membrane-binding region in the dimer. Oligomerization of dimers occurs on another interface of the nucleotide-binding domain, and this allows us to model the EHD oligomer. We discuss the functional implications of the EHD2 structure for understanding membrane deformation.

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Figure 1: EHD2 shares common properties with the dynamin superfamily.
Figure 2: The structure of EHD2.
Figure 3: Membrane binding and the role of ATP hydrolysis.
Figure 4: The EHD2 oligomer.

Change history

  • 18 October 2007

    The AOP version of this paper contained the term 'Epsin homology (EH)-domain...'. This was corrected to 'Eps15 homology (EH)-domain...' in the 18 October issue


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Long-term fellowships supported O.D. (The International Human Frontier Science Program Organization), R.L. (Swedish Research Council) and S.M. (EMBO). We thank M. Plomann for providing the complementary DNAs for mammalian EHDs, and the ESRF beam staff in Grenoble for their support. The authors declare no competing financial interests.

The atomic coordinates of mouse EHD2 have been deposited in the Protein Data Bank (PDB) with the accession number 2QPT.

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Correspondence to Oliver Daumke or Harvey T. McMahon.

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Supplementary information

Supplementary Information 1

This file contains Supplementary Figures 1-12 with Legends, Supplementary Table 1 with the data collection statistics and additional references. (PDF 2225 kb)

Supplementary Information 2

The file contains Supplementary Video 1 which shows EHD2 wild-type was over-expressed in HeLa cells for 24 h and imaged by EPI-fluorescence for approximately 30 min. Some of the tubules and puncta are dynamic. (MOV 4707 kb)

Supplementary Information 3

The file contains Supplementary Video 2 which shows cropped area of EHD2 WT (Video 1) at higher resolution. (MOV 5319 kb)

Supplementary Information 4

The file contains Supplementary Video 3 which shows EHD2 T94A was over-expressed in HeLa cells for 24 h and imaged by EPI-fluorescence for approximately 30 min. There are only tubules, and these are mostly static. Cropped movies show a smaller area at higher resolution. (MOV 6652 kb)

Supplementary Information 5

The file contains Supplementary Video 4 which shows cropped area of EHD2 T94A (Video 2) at higher resolution. (MOV 7726 kb)

Supplementary Information 6

The file contains Supplementary Video 5 which shows EHD2 I157Q was over-expressed in HeLa cells for 24h and imaged by EPI-fluorescence for approximately 30min. No tubules can be found and the puncta are mostly motile. (MOV 9013 kb)

Supplementary Information 7

The file contains Supplementary Video 6 which shows cropped area of EHD2 I157Q (Video 4) at higher resolution. (MOV 6353 kb)

Supplementary Information 8

The file contains Supplementary Data with PDB coordinates of the proposed EHD2 oligomer. Four EHD2 dimers (in the absence of the EH domain) were aligned as described in Methods. All lipid interaction sites point towards the putative membrane interface. Molecules B and C which have been used for the initial alignment with GBP1 are related via a 2-fold axis and the nucleotides of these molecules are oriented in a head-to-head fashion. (ZIP 455 kb)

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Daumke, O., Lundmark, R., Vallis, Y. et al. Architectural and mechanistic insights into an EHD ATPase involved in membrane remodelling. Nature 449, 923–927 (2007).

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