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Structural insights into membrane interaction and caveolar targeting of dynamin-like EHD2

Item Type:Article
Title:Structural insights into membrane interaction and caveolar targeting of dynamin-like EHD2
Creators Name:Shah, C. and Hegde, B.G. and Morén, B. and Behrmann, E. and Mielke, T. and Moenke, G. and Spahn, C.M. and Lundmark, R. and Daumke, O. and Langen, R.
Abstract:The dynamin-related Eps15-homology domain-containing protein 2 (EHD2) is a membrane-remodeling ATPase that regulates the dynamics of caveolae. Here, we established an electron paramagnetic resonance (EPR) approach to characterize structural features of membrane-bound EHD2. We show that residues at the tip of the helical domain can insert into the membrane and may create membrane curvature by a wedging mechanism. Using EPR and X-ray crystallography, we found that the N terminus is folded into a hydrophobic pocket of the GTPase domain in solution and can be released into the membrane. Cryoelectron microscopy demonstrated that the N terminus is not essential for oligomerization of EHD2 into a membrane-anchored scaffold. Instead, we found a function of the N terminus in regulating targeting and stable association of EHD2 to caveolae. Our data uncover an unexpected, membrane-induced regulatory switch in EHD2 and demonstrate the versatility of EPR to study structure and function of dynamin superfamily proteins.
Keywords:3T3-L1 Cells, Binding Sites, Carrier Proteins, Caveolae, Cell Membrane, Cryoelectron Microscopy, Electron Spin Resonance Spectroscopy, Green Fluorescent Proteins, HeLa Cells, Molecular Models, Protein Folding, Protein Stability, Tertiary Protein Structure, X-Ray Crystallography, Animals, Mice
Publisher:Cell Press / Elsevier
Page Range:409-420
Date:4 March 2014
Official Publication:https://doi.org/10.1016/j.str.2013.12.015
PubMed:View item in PubMed

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