Curvature of clathrin-coated pits driven by epsin

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Clathrin-mediated endocytosis involves cargo selection and membrane budding into vesicles with the aid of a protein coat. Formation of invaginated pits on the plasma membrane and subsequent budding of vesicles is an energetically demanding process that involves the cooperation of clathrin with many different proteins. Here we investigate the role of the brain-enriched protein epsin 1 in this process. Epsin is targeted to areas of endocytosis by binding the membrane lipid phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2). We show here that epsin 1 directly modifies membrane curvature on binding to PtdIns(4,5)P2 in conjunction with clathrin polymerization. We have discovered that formation of an amphipathic α-helix in epsin is coupled to PtdIns(4,5)P2 binding. Mutation of residues on the hydrophobic region of this helix abolishes the ability to curve membranes. We propose that this helix is inserted into one leaflet of the lipid bilayer, inducing curvature. On lipid monolayers epsin alone is sufficient to facilitate the formation of clathrin-coated invaginations.

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Figure 1: Epsin 1 ENTH domain tubulates liposomes.
Figure 2: Structure of epsin ENTH bound to Ins(1,4,5)P3.
Figure 3: Binding of epsin ENTH and mutants to phosphoinositides.
Figure 4: Effect of full-length epsin and mutants on the distribution of the AP2 complex in COS-7 cells visualized by sequential imaging with confocal microscopy.
Figure 5: Mutations of L6 on helix 0 affect the ability of epsin to tubulate liposomes.
Figure 6: Clathrin recruitment to lipid monolayers by epsin.


  1. 1

    Cremona, O. & De Camilli, P. Synaptic vesicle endocytosis. Curr. Opin. Neurobiol. 7, 323–330 (1997)

  2. 2

    Schmid, S. L. Clathrin-coated vesicle formation and protein sorting: an integrated process. Annu. Rev. Biochem. 66, 511–548 (1997)

  3. 3

    Hirst, J. & Robinson, M. S. Clathrin and adaptors. Biochim. Biophys. Acta 1404, 173–193 (1998)

  4. 4

    Marsh, M. & McMahon, H. T. The structural era of endocytosis. Science 285, 215–220 (1999)

  5. 5

    Brodsky, F. M., Chen, C. Y., Knuehl, C., Towler, M. C. & Wakeham, D. E. Biological basket weaving: formation and function of clathrin-coated vesicles. Annu. Rev. Cell Dev. Biol. 17, 517–568 (2001)

  6. 6

    Schmidt, A. et al. Endophilin I mediates synaptic vesicle formation by transfer of arachidonate to lysophosphatidic acid. Nature 401, 133–141 (1999)

  7. 7

    Salcini, A. E., Chen, H., Iannolo, G., De Camilli, P. & Di Fiore, P. P. Epidermal growth factor pathway substrate 15, Eps15. Int. J. Biochem. Cell Biol. 31, 805–809 (1999)

  8. 8

    McPherson, P. S. et al. EH domain-dependent interactions between Eps15 and clathrin-coated vesicle protein p95. Biochem. Biophys. Res. Commun. 244, 701–705 (1998)

  9. 9

    Chen, H. et al. Epsin is an EH-domain-binding protein implicated in clathrin-mediated endocytosis. Nature 394, 793–797 (1998)

  10. 10

    Cadavid, A. L., Ginzel, A. & Fischer, J. A. The function of the Drosophila fat facets deubiquitinating enzyme in limiting photoreceptor cell number is intimately associated with endocytosis. Development 127, 1727–1736 (2000)

  11. 11

    Wendland, B., Steece, K. E. & Emr, S. D. Yeast epsins contain an essential N-terminal ENTH domain, bind clathrin and are required for endocytosis. EMBO J. 18, 4383–4393 (1999)

  12. 12

    Itoh, T. et al. Role of the ENTH domain in phosphatidylinositol-4,5-bisphosphate binding and endocytosis. Science 291, 1047–1051 (2001)

  13. 13

    Ford, M. G. J. et al. Simultaneous binding of PtdIns(4,5)P2 and clathrin by AP180 in the nucleation of clathrin lattices on membranes. Science 291, 1051–1055 (2001)

  14. 14

    Hyman, J., Chen, H., Di Fiore, P. P., De Camilli, P. & Brunger, A. T. Epsin 1 undergoes nucleocytosolic shuttling and its eps15 interactor NH(2)-terminal homology (ENTH) domain, structurally similar to Armadillo and HEAT repeats, interacts with the transcription factor promyelocytic leukemia Zn(2) + finger protein (PLZF). J. Cell Biol. 149, 537–546 (2000)

  15. 15

    Morgan, J. R., Prasad, K., Jin, S., Augustine, G. J. & Lafer, E. M. Uncoating of clathrin-coated vesicles in presynaptic terminals: roles for Hsc70 and auxilin. Neuron 32, 289–300 (2001)

  16. 16

    Drake, M. T., Downs, M. A. & Traub, L. M. Epsin binds to clathrin by associating directly with the clathrin-terminal domain. Evidence for cooperative binding through two discrete sites. J. Biol. Chem. 275, 6479–6489 (2000)

  17. 17

    Sweitzer, S. M. & Hinshaw, J. E. Dynamin undergoes a GTP-dependent conformational change causing vesiculation. Cell 93, 1021–1029 (1998)

  18. 18

    Takei, K., Slepnev, V. I., Haucke, V. & De Camilli, P. Functional partnership between amphiphysin and dynamin in clathrin-mediated endocytosis. Nature Cell Biol. 1, 33–39 (1999)

  19. 19

    Farsad, K. et al. Generation of high curvature membranes mediated by direct endophilin bilayer interactions. J. Cell Biol. 155, 193–200 (2001)

  20. 20

    Zhang, B. et al. Synaptic vesicle size and number are regulated by a clathrin adaptor protein required for endocytosis. Neuron 21, 1465–1475 (1998)

  21. 21

    Kalthoff, C., Alves, J., Urbanke, C., Knorr, R. & Ungewickell, E. J. Unusual structural organization of the endocytic proteins AP180 and epsin 1. J. Biol. Chem. 277, 8209–8216 (2001)

  22. 22

    Tebar, F., Sorkina, T., Sorkin, A., Ericsson, M. & Kirchhausen, T. Eps15 is a component of clathrin-coated pits and vesicles and is located at the rim of coated pits. J. Biol. Chem. 271, 28727–28730 (1996)

  23. 23

    Owen, D. J., Vallis, Y., Pearse, B. M., McMahon, H. T. & Evans, P. R. The structure and function of the beta 2-adaptin appendage domain. EMBO J. 19, 4216–4227 (2000)

  24. 24

    Nossal, R. Energetics of clathrin basket assembly. Traffic 2, 138–147 (2001)

  25. 25

    Owen, D. J. et al. A structural explanation for the binding of multiple ligands by the α-adaptin appendage domain. Cell 97, 805–815 (1999)

  26. 26

    Razzaq, A. et al. Amphiphysin is necessary for organization of the excitation-contraction coupling machinery of muscles, but not for synaptic vesicle endocytosis in Drosophila. Genes Dev. 15, 2967–2979 (2001)

  27. 27

    Hopp, T. P. & Woods, K. R. Prediction of protein antigenic determinants from amino acid sequences. Proc. Natl Acad. Sci. USA 78, 3824–3828 (1981)

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We thank O. Perisic and R. Williams for advice on protein–membrane interactions; B. Collins and staff at the ESRF for assistance with data collection; J. Butler for assistance with analytical ultracentrifugation; and J. Berriman for assistance with electron microscopy. We also thank B. Pearse, N. Unwin, M. Stowell, M. Goedert, S. Munro, P. Rosenthal and members of our laboratories for extensive discussions, and J. Gallop for a supply of dynamin. I.G.M. was supported by an MRC Postdoctoral Fellowship, B.J.P was supported by an EMBO Long Term Fellowship, and G.J.K.P. was supported by a Marie Curie Fellowship.

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Correspondence to Philip R. Evans or Harvey T. McMahon.

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Ford, M., Mills, I., Peter, B. et al. Curvature of clathrin-coated pits driven by epsin. Nature 419, 361–366 (2002) doi:10.1038/nature01020

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