Curvature sensing by the epsin N-terminal homology domain measured on cylindrical lipid membrane tethers.
The protein epsin is believed to play important roles in clathrin-mediated endocytosis, including generation of the high membrane curvature necessary for vesicle formation. Here we assess the basis for this hypothesis by systematically quantifying the curvature dependence of the area density of epsin N-terminal homology (ENTH) domain on cylindrical membranes of controlled curvature. In cylindrical tethers pulled from micropipet-aspirated giant unilamellar vesicles, repartitioning of membrane-bound ENTH from vesicles onto highly curved membranes was observed by fluorescence microscopy. First-order thermodynamic theory used to analyze our data yielded the first measurement of Leibler's thermodynamic curvature-composition coupling coefficient to be reported for an endocytic accessory protein. Our results highlight the possibility that epsin contributes to cellular membrane curvature sensing and generation, and we believe that our method will provide useful contributions toward the goal of relating molecular descriptions of interactions to macroscopic membrane remodeling in cells and identifying and characterizing roles for proteins in these processes.
Capraro
Benjamin R
BR
Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Yoon
Youngdae
Y
Cho
Wonhwa
W
Baumgart
Tobias
T
D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013816:Thermodynamics; D053835:Unilamellar Liposomes / Q000378:metabolism*
D033942:Adaptor Proteins, Vesicular Transport; D008563:Membrane Lipids; D053835:Unilamellar Liposomes; C114154:epsin