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Two distinct interaction motifs in amphiphysin bind two independent sites on the clathrin terminal domain β-propeller

Nature Structural & Molecular Biology volume 11, pages 242–248 (2004)Cite this article

Abstract

During the assembly of clathrin-coated vesicles, many peripheral membrane proteins, including the amphiphysins, use LLDLD-type clathrin-box motifs to interact with the N-terminal β-propeller domain (TD) of clathrin. The 2.3 Å–resolution structure of the clathrin TD in complex with a TLPWDLWTT peptide from amphiphysin 1 delineates a second clathrin-binding motif, PWXXW (the W box), that binds at a site on the TD remote from the clathrin box–binding site. The presence of both sequence motifs within the unstructured region of the amphiphysins allows them to bind more tightly to free TDs than do other endocytic proteins that contain only clathrin-box motifs. This property, along with the propensity of the N-terminal BAR domain to bind curved membranes, will preferentially localize amphiphysin and its partner, dynamin, to the periphery of invaginated clathrin lattices.

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Figure 1: Overall structure of the bovine clathrin TD in complex with the W-box peptide (TLPWDLWTT) from human amphiphysin 1.
Figure 2: Molecular detail of the W-box peptide-binding site.
Figure 3: Amphiphysin-TD interactions.
Figure 5: Interaction of amphiphysin and SNX9 with cytosolic clathrin trimers.
Figure 4: Schematic representation of clathrin TD bound to the insert domain of amphiphysin that contains both a clathrin-box and a W-box motif.

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Acknowledgements

We thank the staff of Daresbury Synchrotron Radiation Source and M. Ford for help in data collection, and the Wellcome Trust for a traveling research fellowship to A.E.M. and for a senior research fellowship in basic biomedical sciences to D.J.O. L.M.T. was supported in part by US National Institutes of Health grant R01 DK53249.

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  1. Adriana E Miele

    Present address: Department of Biochemical Sciences, University of Rome La Sapienza, Piazzale Aldo Moro 5, 00185, Rome, Italy

Authors and Affiliations

  1. MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK

    Adriana E Miele & Philip R Evans

  2. Cambridge Institute for Medical Research, University of Cambridge, Wellcome Trust/MRC building, Hills Road, Cambridge, CB2 2XY, UK

    Peter J Watson & David J Owen

  3. Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, 15261, Pennsylvania, USA

    Linton M Traub

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Correspondence to Linton M Traub or David J Owen.

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Miele, A., Watson, P., Evans, P. et al. Two distinct interaction motifs in amphiphysin bind two independent sites on the clathrin terminal domain β-propeller. Nat Struct Mol Biol 11, 242–248 (2004). https://doi.org/10.1038/nsmb736

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