title: Spatial Control of Epsin-induced Clathrin Assembly by Membrane Curvature.
abstract: Epsins belong to the family of highly conserved clathrin-associated sorting proteins that are indispensable for clathrin-mediated endocytosis, but their precise functions remain unclear. We have developed an assay system of budded supported membrane tubes displaying planar and highly curved membrane surfaces to analyze intrinsic membrane curvature preference shown by clathrin-associated sorting proteins. Using real-time fluorescence microscopy, we find that epsin preferentially partitions to and assembles clathrin on highly curved membrane surfaces. Sorting of epsin to regions of high curvature strictly depends on binding to phosphatidylinositol 4,5-bisphosphate. Fluorescently labeled clathrins rapidly assemble as foci, which in turn cluster epsin, while maintaining tube integrity. Clathrin foci grow in intensity with a typical time constant of ∼75 s, similar to the time scales for coated pit formation seen in cells. Epsin therefore effectively senses membrane curvature to spatially control clathrin assembly. Our results highlight the potential role of membrane curvature in orchestrating the myriad molecular interactions necessary for the success of clathrin-mediated membrane budding.
authors.affiliation: From the Indian Institute of Science Education and Research, Dr. Homi Bhabha Road, Pashan, Pune 411 008, India.
authors.forename: Sachin S
authors.initials: SS
authors.lastname: Holkar
authors.affiliation: From the Indian Institute of Science Education and Research, Dr. Homi Bhabha Road, Pashan, Pune 411 008, India.
authors.forename: Sukrut C
authors.initials: SC
authors.lastname: Kamerkar
authors.affiliation: From the Indian Institute of Science Education and Research, Dr. Homi Bhabha Road, Pashan, Pune 411 008, India pucadyil@iiserpune.ac.in.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Pucadyil
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002966:Clathrin / Q000737:chemistry / Q000378:metabolism*; D004705:Endocytosis; D006041:Goats; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D051381:Rats
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D002966:Clathrin; D008567:Membranes, Artificial; C114154:epsin
keywords: clathrin; endocytosis; epsin; fluorescence; membrane curvature sensing; membrane structure; microscopy
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