Structural Basis of Membrane Bending by the N-BAR Protein Endophilin

Department of Molecular Biosciences, Northwestern University, 2205 Campus Drive, Evanston, IL 60208, USA.
Cell (Impact Factor: 32.24). 03/2012; 149(1):137-45. DOI: 10.1016/j.cell.2012.01.048
Source: PubMed


Functioning as key players in cellular regulation of membrane curvature, BAR domain proteins bend bilayers and recruit interaction partners through poorly understood mechanisms. Using electron cryomicroscopy, we present reconstructions of full-length endophilin and its N-terminal N-BAR domain in their membrane-bound state. Endophilin lattices expose large areas of membrane surface and are held together by promiscuous interactions between endophilin's amphipathic N-terminal helices. Coarse-grained molecular dynamics simulations reveal that endophilin lattices are highly dynamic and that the N-terminal helices are required for formation of a stable and regular scaffold. Furthermore, endophilin accommodates different curvatures through a quantized addition or removal of endophilin dimers, which in some cases causes dimerization of endophilin's SH3 domains, suggesting that the spatial presentation of SH3 domains, rather than affinity, governs the recruitment of downstream interaction partners.

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    ABSTRACT: The generation of neuronal morphology requires transport vesicles originating from the Golgi apparatus (GA) to deliver specialized components to the axon and dendrites. Drosophila Arfaptin is a membrane-binding protein localized to the GA that is required for the growth of the presynaptic nerve terminal. Here we provide biochemical, cellular and genetic evidence that the small GTPase Arl1 and the guanine-nucleotide exchange factor (GEF) Gartenzwerg are required for Arfaptin function at the Golgi during synapse growth. Our data define a new signaling pathway composed of Arfaptin, Arl1, and Garz, required for the generation of normal synapse morphology. © 2015. Published by The Company of Biologists Ltd.
    Biology Open 06/2015; 4(8). DOI:10.1242/bio.011262 · 2.42 Impact Factor
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    • "The helix mode based on the arrangement of endophilin BAR domains on tubules (Mim et al., 2012) also yielded good fits to the experimental data, although inferior to the fits from the offset mode (Figure 6B). On the other hand, the tip-to-tip, side-byside , and circle mode produced substantially worse fits (Figures 10Å A "
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    ABSTRACT: PICK1 is a neuronal scaffolding protein containing a PDZ domain and an auto-inhibited BAR domain. BAR domains are membrane-sculpting protein modules generating membrane curvature and promoting membrane fission. Previous data suggest that BAR domains are organized in lattice-like arrangements when stabilizing membranes but little is known about structural organization of BAR domains in solution. Through a small-angle X-ray scattering (SAXS) analysis, we determine the structure of dimeric and tetrameric complexes of PICK1 in solution. SAXS and biochemical data reveal a strong propensity of PICK1 to form higher-order structures, and SAXS analysis suggests an offset, parallel mode of BAR-BAR oligomerization. Furthermore, unlike accessory domains in other BAR domain proteins, the positioning of the PDZ domains is flexible, enabling PICK1 to perform long-range, dynamic scaffolding of membrane-associated proteins. Together with functional data, these structural findings are compatible with a model in which oligomerization governs auto-inhibition of BAR domain function. Copyright © 2015 Elsevier Ltd. All rights reserved.
    Structure 06/2015; 23(7). DOI:10.1016/j.str.2015.04.020 · 5.62 Impact Factor
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    • "It is, however, also important to note that the self-assembly mechanisms of N-BAR and F-BAR domains are different. Protein scaffolds formed by endophilin N-BAR domain are held together through interactions between endophilin's amphipathic N-terminal helices, whereas the F-BAR domain scaffolds are stabilized through lateral contacts between the coiled-coil regions of the domains (Frost et al., 2008; Mim et al., 2012). Importantly, membrane insertion of the N-terminal a helix of Rvs161/167, endophilin, and amphiphysin BAR domains is enhanced by PI(4,5)P 2 (Figures S3E and S3F; Yoon et al., 2012), demonstrating that, in addition to the coiled-coil region, also the N-terminal helix contributes to phosphoinositide specificity of N- BAR domains. "
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    ABSTRACT: Bin-Amphiphysin-Rvs (BAR) domain proteins are central regulators of many cellular processes involving membrane dynamics. BAR domains sculpt phosphoinositide-rich membranes to generate membrane protrusions or invaginations. Here, we report that, in addition to regulating membrane geometry, BAR domains can generate extremely stable lipid microdomains by "freezing" phosphoinositide dynamics. This is a general feature of BAR domains, because the yeast endocytic BAR and Fes/CIP4 homology BAR (F-BAR) domains, the inverse BAR domain of Pinkbar, and the eisosomal BAR protein Lsp1 induced phosphoinositide clustering and halted lipid diffusion, despite differences in mechanisms of membrane interactions. Lsp1 displays comparable low diffusion rates in vitro and in vivo, suggesting that BAR domain proteins also generate stable phosphoinositide microdomains in cells. These results uncover a conserved role for BAR superfamily proteins in regulating lipid dynamics within membranes. Stable microdomains induced by BAR domain scaffolds and specific lipids can generate phase boundaries and diffusion barriers, which may have profound impacts on diverse cellular processes.
    Cell Reports 09/2013; 4(6). DOI:10.1016/j.celrep.2013.08.024 · 8.36 Impact Factor
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