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Clathrin self-assembly is mediated by a tandemly repeated superhelix

Nature volume 399, pages 371–375 (1999)Cite this article

Abstract

Clathrin is a triskelion-shaped cytoplasmic protein that polymerizes into a polyhedral lattice on intracellular membranes to form protein-coated membrane vesicles. Lattice formation induces the sorting of membrane proteins during endocytosis and organelle biogenesis by interacting with membrane-associated adaptor molecules1. The clathrin triskelion is a trimer of heavy-chain subunits (1,675 residues), each binding a single light-chain subunit, in the hub domain (residues 1,074–1,675). Light chains negatively modulate polymerization so that intracellular clathrin assembly is adaptor-dependent2. Here we report the atomic structure, to 2.6 Å resolution, of hub residues 1,210–1,516 involved in mediating spontaneous clathrin heavy-chain polymerization and light-chain association3,4. The hub fragment folds into an elongated coil of α-helices, and alignment analyses reveal a 145-residue motif that is repeated seven times along the filamentous leg and appears in other proteins involved in vacuolar protein sorting. The resulting model provides a three-dimensional framework for understanding clathrin heavy-chain self-assembly, light-chain binding and trimerization.

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Figure 1: Organization of the clathrin polyhedron and triskelion.
Figure 2: The crystal structure of clathrin heavy-chain residues 1,210–1,516.
Figure 3: Optimized superposition of Cα traces from CHCR5, CHCR6 and CHCR7 regions in the clathrin proximal leg, determined with LSQMAN27.
Figure 4: Sequence analysis of clathrin heavy-chain and related sequences.
Figure 5: Surface representations of clathrin heavy chain, residues 1210–1516.

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Acknowledgements

We thank F. Masiarz for mass spectrometry analysis; L.-W. Hung for assistance with data collection at beamline 5.0.2 of the Macromolecular Cyrstallography Facility at the Advanced Light Source (ALS is funded by the U.S. Department of Energy Office of Basic Energy Sciences); H. Bellamy and W. Weis for discussions on crystallographic data collection and analysis; T. Terwilliger and C. Weekes for correspondence on SOLVE and SnB2, respectively; M. Butte, for software assistance and comments on manuscript. This work was supported by the NIH (F.M.B., R.J.F.)

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Author notes

  1. Kai Lin

    Present address: Department of Pharmacology, University of Massachusetts Medical Center, 55 Lake Ave., North Worcester, Massachusetts, 01655, USA

Authors and Affiliations

  1. The Department of Microbiology and Immunology, Departments of Biopharmaceutical Sciences and Pharmaceutical Chemistry, G. W. Hooper Foundation,

    Joel A. Ybe, Frances M. Brodsky, Shu-Hui Liu & Lin Chen

  2. Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, 94143, California, USA

    Kai Lin, Robert J. Fletterick & Peter K. Hwang

  3. Bioinformatics Group, MEMOREC Stoffel GmbH, Stoeckheimer Weg 1, Koeln, 50829, Germany

    Kay Hofmann

  4. Physical Biosciences Division, Macromolecular Cyrstallography Facility at the Advanced Light Source, Lawrence Berkeley National Laboraotry, Berkeley, 94720, California, USA

    Thomas N. Earnest

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Correspondence to Frances M. Brodsky.

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Ybe, J., Brodsky, F., Hofmann, K. et al. Clathrin self-assembly is mediated by a tandemly repeated superhelix. Nature 399, 371–375 (1999). https://doi.org/10.1038/20708

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