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Epsin is an EH-domain-binding protein implicated in clathrin-mediated endocytosis

Nature volume 394, pages 793–797 (1998)Cite this article

Abstract

During endocytosis, clathrin and the clathrin adaptor protein AP-2 (ref. 1), assisted by a variety of accessory factors, help to generate an invaginated bud at the cell membrane2,3. One of these factors is Eps15, a clathrin-coat-associated protein that binds the α-adaptin subunit of AP-2 (4–8). Here we investigate the function of Eps15 by characterizing an important binding partner for its region containing EH domains9; this protein, epsin, is closely related to the Xenopus mitotic phosphoprotein MP90 (ref. 10) and has a ubiquitous tissue distribution. It is concentrated together with Eps15 in presynaptic nerve terminals, which are sites specialized for the clathrin-mediated endocytosis of synaptic vesicles. The central region of epsin binds AP-2 and its carboxy-terminal region binds Eps15. Epsin is associated with clathrin coats in situ, can be co-precipitated with AP-2 and Eps15 from brain extracts, but does not co-purify with clathrin coat components in a clathrin-coated vesicle fraction. When epsin function is disrupted, clathrin-mediated endocytosis is blocked. We propose that epsin may participate, together with Eps15, in the molecular rearrangement of the clathrin coats that are required for coated-pit invagination and vesicle fission.

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Figure 1: Eps15 and epsin are highly expressed in the brain and are concentrated in nerve terminals.
Figure 2: EH-domain-containing regions of Eps15 and Eps15R bind to a 94K protein (epsin) in rat brain.
Figure 3: Primary sequence of rat epsin.
Figure 4: Subcellular localization of epsin.
Figure 5: Interaction of epsin with α-adaptin and Eps15.
Figure 6: Perturbation of epsin function impairs receptor-mediated endocytosis.

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Acknowledgements

We thank Y. Nemoto for help in preliminary experiments, R. Bauerfeind and O.Cremona for advice, L. Daniell and M. Salazar for help with electron microscopy, X. Zhang for technical assistance and support, and M. M. Zhou for discussion. This work was supported in part by grants from the NIH, the HFSP and the US Army Medical Research and Development Command (to P.D.C.), and from the Associazione Italiana Ricerca sul Cancro, the Consiglio Nazionale delle Ricerche, the European Community (BIOMED-2 Programme), the Armenise-Harvard Foundation and the Ferrero Foundation (to P.P.D.F.).

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Authors and Affiliations

  1. Howard Hughes Medical Institute and Department of Cell Biology, Yale University School of Medicine, 295 Congress Avenue, New Haven, 06510, Connecticut, USA

    Hong Chen, Vladimir I. Slepnev, Kohji Takei, Margaret H. Butler & Pietro De Camilli

  2. Department of Experimental Oncology, European Institute of Oncology, Milan, 20141, Italy

    Silvia Fre, Maria Rosaria Capua & Pier Paolo Di Fiore

  3. Istituto di Microbiologia, Universita' di Bari, Bari, 70124, Italy

    Pier Paolo Di Fiore

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Correspondence to Pietro De Camilli.

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Chen, H., Fre, S., Slepnev, V. et al. Epsin is an EH-domain-binding protein implicated in clathrin-mediated endocytosis. Nature 394, 793–797 (1998). https://doi.org/10.1038/29555

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