Molecular Basis of the Potent Membrane-remodeling Activity of the Epsin 1 N-terminal Homology Domain

Yoon, Yonghan; Jiao, Tong; Lee, Park Joo; Albanese, Alexandra; Bhardwaj, Nitin; Källberg, Morten; Digman, Michelle A.; Lu, Hui; Gratton, Enrico; Shin, Young Kil; Cho, Wonhwa

doi:10.1074/jbc.m109.068015

Cited by 60 publications

(80 citation statements)

References 46 publications

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“…This fact helps explain the remodeling activity of epsin proteins (9,41), which exhibit no apparent special shape, and provides insights into the mechanics of tubulation induced by crowding of proteins and anchored polymers (15,42). We also demonstrated that regardless of the orientation of their N-terminal helices, the proteins may assemble rapidly to induce membrane buds.…”

Section: Discussionmentioning

confidence: 89%

Linear aggregation of proteins on the membrane as a prelude to membrane remodeling

Šimunović

Srivastava

Voth

2013

Proc. Natl. Acad. Sci. U.S.A.

143

183

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Adhesion and insertion of curvature-mediating proteins can induce dramatic structural changes in cell membranes, allowing them to participate in several key cellular tasks. The way proteins interact to generate curvature remains largely unclear, especially at early stages of membrane remodeling. Using a coarse-grained model of Bin/ amphiphysin/Rvs domain with an N-terminal helix (N-BAR) interacting with flat membranes and vesicles, we demonstrate that at low protein surface densities, binding of N-BAR domain proteins to the membrane is followed by a linear aggregation and the formation of meshes on the surface. In this process, the proteins assemble at the base of emerging membrane buds. Our work shows that beyond a more straightforward scaffolding mechanism at high bound densities, the interplay of anisotropic interactions and the local stress imposed by the N-BAR proteins results in deep invaginations and endocytic vesicular bud-like deformations, an order of magnitude larger than the size of the individual protein. Our results imply that by virtue of this mechanism, cell membranes may achieve rapid local increases in protein concentration.coarse-grained simulation | membrane stress | self-assembly | membrane curvature | Gaussian curvature L ipid membranes protect cells and their organelles and serve as mechanical support for proteins involved in signal transduction and cellular trafficking (1). Owing to the way lipids are assembled into bilayers of mesoscopic length and molecular width, membranes behave as elastic and highly dynamic molecular sheets. Their innate multiscale nature is further evident in the local interplay between proteins and lipids, which results in large-scale membrane remodeling characterized by an impressive array of morphologies (2). These properties allow biological membranes to take part in several important cellular processes, as their restructuring is key to enabling communication between cells, formation of organelles, trafficking, division, and cell migration (1).The interactions between individual proteins and lipid molecules alone are insufficient to remodel flat membrane sheets to an appreciable extent, implying that the experimentally observed membrane remodeling is a result of the concerted action of multiple proteins (3) in ways mechanistically differing from the mode of action of a single protein. Considering the multiple timescales required to capture protein dynamics as the membrane deformations are induced, as well as the difficulty in separating individual events of the remodeling process, this mechanism remains largely unclear. It previously was demonstrated that the binding of curvature-inducing nanoparticles may form tubular and vesicular structures, in which particles interact with one another solely via curvature (3-6). Furthermore, analytical theory predicts that anisotropic inclusions may induce budding on the membrane surface (7). However, the way proteins assemble on the membrane and how their oligomerization couples to both local and long-wavelength transformat...

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Section: Discussionmentioning

confidence: 89%

Linear aggregation of proteins on the membrane as a prelude to membrane remodeling

Šimunović

Srivastava

Voth

2013

Proc. Natl. Acad. Sci. U.S.A.

143

183

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“…It has been shown that upon binding of ENTH to PtdIns(4,5)P 2 , the protein experiences a change in structure in which helix-0 inserts into the lipid bilayer as demonstrated by EPR spectroscopy (22). This increases the surface area of the outer leaflet of the GUV membrane and induces an asymmetry in the two bilayer leaflets facilitating membrane bending by generating spontaneous curvature (26,45).…”

Section: ϫ4mentioning

confidence: 99%

“…ENTH Binding to PtdIns(4,5)P 2 -doped GUVs-ENTH can induce membrane tubulation (22,25). Recently we showed that binding of ENTH to PtdIns(4,5)P 2 -doped and slightly curved pore-spanning membranes results in a reduction of the lateral membrane tension (27).…”

Section: ϫ4mentioning

confidence: 99%

Epsin N-terminal Homology Domain (ENTH) Activity as a Function of Membrane Tension

Gleisner

Kroppen

Fricke

et al. 2016

Journal of Biological Chemistry

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The epsin N-terminal homology domain (ENTH) is a major player in clathrin-mediated endocytosis. To investigate the influence of initial membrane tension on ENTH binding and activity, we established a bilayer system based on adhered giant unilamellar vesicles (GUVs) to be able to control and adjust the membrane tension covering a broad regime. The shape of each individual adhered GUV as well as its adhesion area was monitored by spinning disc confocal laser microscopy. Control of in a range of 0.08-1.02 mN/m was achieved by altering the Mg 2؉ concentration in solution, which changes the surface adhesion energy per unit area of the GUVs. Specific binding of ENTH to phosphatidylinositol 4,5-bisphosphate leads to a substantial increase in adhesion area of the sessile GUV. At low tension (<0.1 mN/m) binding of ENTH can induce tubular structures, whereas at higher membrane tension the ENTH interaction deflates the sessile GUV and thereby increases the adhesion area. The increase in adhesion area is mainly attributed to a decrease in the area compressibility modulus K A . We propose that the insertion of the ENTH helix-0 into the membrane is largely responsible for the observed decrease in K A , which is supported by the observation that the mutant ENTH L6E shows a reduced increase in adhesion area. These results demonstrate that even in the absence of tubule formation, the area compressibility modulus and, as such, the bending rigidity of the membrane is considerably reduced upon ENTH binding. This renders membrane bending and tubule formation energetically less costly.Clathrin-mediated endocytosis is one of the key metabolic pathways for the uptake of macromolecules into eukaryotic cells (1-4). Driven by a chain of remodeling events and an elaborate set of proteins acting in an orchestrated manner, an almost flat patch of plasma membrane is transformed into a closed, cargo-containing vesicle. As plasma membrane shape transformation is associated with significant local bending of the membrane, the process is highly sensitive to lateral membrane tension. Plasma membrane tension originates from two primary sources; that is, hydrostatic pressure across the lipid bilayer and cytoskeleton-membrane adhesion (5). Depending on the cell type, plasma membrane tensions span a range of roughly 0.003-0.45 mN/m (5-8). Even though it had become clear already in the late 1990s that tension plays a role in exoand endocytosis (9, 10), only in recent years has significant evidence been accumulated that membrane tension is of utmost importance for processes that rely on membrane remodeling (11-14). Cells actively maintain and regulate their membrane tension and use it to control exo-and endocytosis (15). In K562 cells it has been reported that endocytosis is completely suppressed under hypoosmotic conditions (16). Generally, high lateral membrane tension suppresses membrane deformation as both stretching the lipid bilayer and opening bonds between the cytoskeleton and the plasma membrane requires a large amount of energy (17).One protein...

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“…Sla2 and HIP1R form dimers via their coiled-coil domains (14,34), and the ENTH domain of mammalian epsin may oligomerize on liposomes in vitro (35). In addition, both proteins bind clathrin and other coat proteins (reviewed in ref.…”

Section: Sla2∆thatch-acb-gfp Ent1∆acbmentioning

confidence: 99%

Molecular basis for coupling the plasma membrane to the actin cytoskeleton during clathrin-mediated endocytosis

Skružný

Brach

Ciuffa

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

136

203

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Dynamic actin filaments are a crucial component of clathrin-mediated endocytosis when endocytic proteins cannot supply enough energy for vesicle budding. Actin cytoskeleton is thought to provide force for membrane invagination or vesicle scission, but how this force is transmitted to the plasma membrane is not understood. Here we describe the molecular mechanism of plasma membrane-actin cytoskeleton coupling mediated by cooperative action of epsin Ent1 and the HIP1R homolog Sla2 in yeast Saccharomyces cerevisiae. Sla2 anchors Ent1 to a stable endocytic coat by an unforeseen interaction between Sla2's ANTH and Ent1's ENTH lipid-binding domains. The ANTH and ENTH domains bind each other in a liganddependent manner to provide critical anchoring of both proteins to the membrane. The C-terminal parts of Ent1 and Sla2 bind redundantly to actin filaments via a previously unknown phospho-regulated actin-binding domain in Ent1 and the THATCH domain in Sla2. By the synergistic binding to the membrane and redundant interaction with actin, Ent1 and Sla2 form an essential molecular linker that transmits the force generated by the actin cytoskeleton to the plasma membrane, leading to membrane invagination and vesicle budding.ANTH domain | membrane remodeling | PIP 2

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Molecular Basis of the Potent Membrane-remodeling Activity of the Epsin 1 N-terminal Homology Domain

Cited by 60 publications

References 46 publications

Linear aggregation of proteins on the membrane as a prelude to membrane remodeling

Linear aggregation of proteins on the membrane as a prelude to membrane remodeling

Epsin N-terminal Homology Domain (ENTH) Activity as a Function of Membrane Tension

Molecular basis for coupling the plasma membrane to the actin cytoskeleton during clathrin-mediated endocytosis

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