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Re-engineering protein interfaces yields copper-inducible ferritin cage assembly

Nature Chemical Biology volume 9, pages 169–176 (2013)Cite this article

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Abstract

The ability to chemically control protein-protein interactions would allow the interrogation of dynamic cellular processes and lead to a better understanding and exploitation of self-assembling protein architectures. Here we introduce a new engineering strategy—reverse metal-templated interface redesign (rMeTIR)—that transforms a natural protein-protein interface into one that only engages in selective response to a metal ion. We have applied rMeTIR to render the self-assembly of the cage-like protein ferritin controllable by divalent copper binding, which has allowed the study of the structure and stability of the isolated ferritin monomer, the demonstration of the primary role of conserved hydrogen-bonding interactions in providing geometric specificity for cage assembly and the uniform chemical modification of the cage interior under physiological conditions. Notably, copper acts as a structural template for ferritin assembly in a manner that is highly reminiscent of RNA sequences that template virus capsid formation.

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Figure 1: Comparative schemes for MeTIR and rMeTIR.
Figure 2: The ferritin cage architecture and intersubunit interactions in the C2 interface.
Figure 3: Solution assembly properties of MIC1.
Figure 4: Key intersubunit interactions in the C2 interface of the MIC1 cage in the presence and absence of copper.
Figure 5: Secondary structure and stability of MIC1.
Figure 6: The structure of dansyl-modified Cys53MIC1 cage.

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Acknowledgements

This work was supported by the US Department of Energy (Division of Materials Sciences, Office of Basic Energy Sciences, Award DE-FG02-10ER46677 to F.A.T.). F.A.T. was additionally supported by the Beckman Foundation, the Sloan Foundation and the National Science Foundation (CHE-0908115). Portions of this research were carried out at the Stanford Synchrotron Radiation Lightsource, operated by Stanford University on behalf of the Department of Energy. We thank J. Brodin for experimental assistance with TEM measurements and J. Faraone Mennella for illustrations.

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  1. Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California, USA.,

    Dustin J E Huard, Kathleen M Kane & F Akif Tezcan

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D.J.E.H. designed and performed experiments, analyzed data and co-wrote the paper. K.M.K. performed experiments. F.A.T. conceived and directed the project, analyzed data and wrote the paper.

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Correspondence to F Akif Tezcan.

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Huard, D., Kane, K. & Tezcan, F. Re-engineering protein interfaces yields copper-inducible ferritin cage assembly. Nat Chem Biol 9, 169–176 (2013). https://doi.org/10.1038/nchembio.1163

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