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Strong underwater adhesives made by self-assembling multi-protein nanofibres

Nature Nanotechnology volume 9, pages 858–866 (2014)Cite this article

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Abstract

Many natural underwater adhesives harness hierarchically assembled amyloid nanostructures to achieve strong and robust interfacial adhesion under dynamic and turbulent environments. Despite recent advances, our understanding of the molecular design, self-assembly and structure–function relationships of these natural amyloid fibres remains limited. Thus, designing biomimetic amyloid-based adhesives remains challenging. Here, we report strong and multi-functional underwater adhesives obtained from fusing mussel foot proteins (Mfps) of Mytilus galloprovincialis with CsgA proteins, the major subunit of Escherichia coli amyloid curli fibres. These hybrid molecular materials hierarchically self-assemble into higher-order structures, in which, according to molecular dynamics simulations, disordered adhesive Mfp domains are exposed on the exterior of amyloid cores formed by CsgA. Our fibres have an underwater adhesion energy approaching 20.9 mJ m−2, which is 1.5 times greater than the maximum of bio-inspired and bio-derived protein-based underwater adhesives reported thus far. Moreover, they outperform Mfps or curli fibres taken on their own and exhibit better tolerance to auto-oxidation than Mfps at pH ≥ 7.0.

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Figure 1: Combinatorial and modular genetic strategy for engineering self-assembling underwater adhesives.
Figure 2: Comparison of monomer, individual fibril and co-assembled fibril structures before and after molecular dynamics simulations for modified CsgA-Mfp3, Mfp5-CsgA and (CsgA-Mfp3)-co-(Mfp5-CsgA) copolymer constructs.
Figure 3: Purification, in vitro self-assembly and characterization of CsgA, CsgA-Mfp3, Mfp5-CsgA and (CsgA-Mfp3)-co-(Mfp5-CsgA) fibres.
Figure 4: Intrinsic fluorescence of CsgA, CsgA-Mfp3, Mfp5-CsgA and (CsgA-Mfp3)-co-(Mfp5-CsgA) copolymer fibres.
Figure 5: Adhesion force measurements and adhesion stability of hybrid adhesive fibres determined by the AFM colloidal probe technique.
Figure 6: Comparison of adhesion performances of different functionalized adhesive fibres with different AFM tips.

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Acknowledgements

The authors thank A. Schwartzman for help in applying AFM colloid nanoparticle technology for measuring adhesive forces and H. Tavakoli Nia (Ortiz group, MIT) for initial discussions regarding this technology. The authors acknowledge help from the NERCE Biomolecule Production Laboratory (Harvard University) for producing part of the cell pellets for protein purification. The authors also thank the Whitehead Institute and the Biopolymers Laboratory in the David H. Koch Institute for Integrated Cancer Research and the Institute for Soldier Nanotechnologies for access to characterization equipment. This research was primarily supported by the Office of Naval Research (N000141310647). This work was also supported in part by the MRSEC Program of the National Science Foundation under award no. DMR-0819762. T.K.L. acknowledges support from the NIH New Innovator Award (1DP2OD008435). The molecular dynamics modelling used the computer cluster and corresponding materials based upon work supported by the National Science Foundation under award no. 0821391.

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Author notes

  1. Chao Zhong

    Present address: Present address: School of Physical Science and Technology, ShanghaiTech University, Shanghai 201210, China,

Authors and Affiliations

  1. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, 02139-4307, Massachusetts, USA

    Chao Zhong, Thomas Gurry, Allen A. Cheng, Zhengtao Deng, Collin M. Stultz & Timothy K. Lu

  2. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, 02139-4307, Massachusetts, USA

    Chao Zhong, Allen A. Cheng, Jordan Downey, Zhengtao Deng & Timothy K. Lu

  3. Synthetic Biology Group, Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, 02139-4307, Massachusetts, USA

    Chao Zhong, Allen A. Cheng, Jordan Downey, Zhengtao Deng & Timothy K. Lu

  4. Computational and Systems Biology Initiative, Massachusetts Institute of Technology, Cambridge, 02139-4307, Massachusetts, USA

    Thomas Gurry, Collin M. Stultz & Timothy K. Lu

  5. The Institute of Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, 02139-4307, Massachusetts, USA

    Collin M. Stultz

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Contributions

T.K.L. directed the research. C.Z. conceived the technical details and designed the experiments. C.Z. performed or participated in all the experiments. J.D. performed experiments in protein expression and purification. Z.D. assisted in collecting and analysing the fluorescence emission and excitation spectra. A.C. constructed the genes. C.M.S. and T.G. designed the simulations. T.G. performed the simulations. C.Z. and T.K.L. wrote the manuscript with help from all authors. All authors contributed to revising the manuscript.

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Correspondence to Timothy K. Lu.

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T.K.L. and C.Z. have filed a patent disclosure with the MIT Technology Licensing Office on this work.

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Zhong, C., Gurry, T., Cheng, A. et al. Strong underwater adhesives made by self-assembling multi-protein nanofibres. Nature Nanotech 9, 858–866 (2014). https://doi.org/10.1038/nnano.2014.199

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