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Durable protein lattices of clathrin that can be functionalized with nanoparticles and active biomolecules

Nature Nanotechnology volume 10, pages 954–957 (2015)Cite this article

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Abstract

Biological molecules that self-assemble and interact with other molecules are attractive building blocks for engineering biological devices. DNA has been widely used for the creation of nanomaterials1, but the use of proteins remains largely unexplored. Here, we show that clathrin can form homogeneous and extended two-dimensional lattices on a variety of substrates, including glass, metal, carbon and plastic. Clathrin is a three-legged protein complex with unique self-assembling properties and is relevant in the formation of membrane transport vesicles in eukaryotic cells2,3. We used a fragment of the adaptor protein epsin to immobilize clathrin lattices on the substrates. The lattices span multiple square millimetres with a regular periodicity of 30 nm and can be functionalized via modified subunits of clathrin with either inorganic nanoparticles or active enzymes. The lattices can be stored for months after crosslinking and stabilization with uranyl acetate. They could be dehydrated and rehydrated without loss of function, offering potential applications in sensing and as biosynthetic reactors.

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Figure 1: Assembly of clathrin lattices on different surfaces.
Figure 2: Regeneration of clathrin lattices.
Figure 3: Recovery of the epsin layer and of the complete lattice after dehydration.
Figure 4: Lattice functionalization with nanoparticles.
Figure 5: Lattice functionalization with enzymes.

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Acknowledgements

The authors thank H. Ungewickell, G. Preiss and C. Lemke for expert technical assistance, the MHH core facility ‘Confocal Laser Microscopy’ for instrumental support and M. Breyvogel for the CAD model. The AFM instrumentation was funded through the ‘Cluster of Excellence and DFG Research Centre Nanoscale Microscopy and Molecular Physiology of the Brain’. M.P. thanks the Göttingen Graduate School for Neurosciences, Biophysics and Molecular Biosciences (GGNB) for a scholarship. All authors are grateful for the support of E.J. Ungewickell regarding protein purification, ideas and discussions.

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Author notes

  1. P. N. Dannhauser and M. Platen: These authors contributed equally to this work

Authors and Affiliations

  1. Institute of Cell Biology, Centre of Anatomy, Hannover Medical School, Hannover, 30625, Germany

    P. N. Dannhauser & H. Böning

  2. Division of Biosciences, Structural & Molecular Biology, University College London, London, WC1E 6BT, UK

    P. N. Dannhauser

  3. IIIrd Institute of Physics, Georg August University, Göttingen, 37073, Germany

    M. Platen & I. A. T. Schaap

  4. Centre for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Göttingen, 37077, Germany

    I. A. T. Schaap

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  1. P. N. Dannhauser
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Contributions

P.N.D. and H.B. prepared the samples. P.N.D. performed the electron and fluorescence microscopy experiments and analysed data. M.P. performed the AFM experiments and analysed data. P.N.D., M.P. and I.A.T.S. designed the experiments and wrote the manuscript.

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Correspondence to P. N. Dannhauser.

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Competing interests

A patent application has been filed by P.N. Dannhauser and E.J. Ungewickell (EP2812029 A1) that includes aspects of the procedure used for the coating of surfaces. So far, the application has exclusively proved to be of scientific value, with no assignable monetary value.

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Dannhauser, P., Platen, M., Böning, H. et al. Durable protein lattices of clathrin that can be functionalized with nanoparticles and active biomolecules. Nature Nanotech 10, 954–957 (2015). https://doi.org/10.1038/nnano.2015.206

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