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Protein camouflage in cytochrome c–calixarene complexes

Nature Chemistry volume 4, pages 527–533 (2012)Cite this article

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Abstract

Small molecules that recognize protein surfaces are important tools for modifying protein interaction properties. Since the 1980s, several thousand studies concerning calixarenes and host–guest interactions have been published. Although there is growing interest in protein–calixarene interactions, only limited structural information has been available to date. We now report the crystal structure of a protein–calixarene complex. The water-soluble p-sulfonatocalix[4]arene is shown to bind the lysine-rich cytochrome c at three different sites. Binding curves obtained from NMR titrations reveal an interaction process that involves two or more binding sites. Together, the data indicate a dynamic complex in which the calixarene explores the surface of cytochrome c. In addition to providing valuable information on protein recognition, the data also indicate that the calixarene is a mediator of protein–protein interactions, with potential applications in generating assemblies and promoting crystallization.

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Figure 1: NMR spectroscopic characterization of the sclx4 binding surface on cytochrome c.
Figure 2: Binding isotherms for the cytochrome c–sclx4 interaction determined by NMR spectroscopy.
Figure 3: Crystal structure of the cytochrome c–sclx4 complex.
Figure 4: Sclx4 explores and camouflages the surface of cytochrome c.

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Acknowledgements

This research was supported by NUI Galway (college scholarship to R.E.M., Millennium Fund grants to N.P. and P.B.C.) and Science Foundation Ireland (grants 07/IN.1/B975 to A.R.K. and 07/RFP/BICF236 to P.B.C.). The authors acknowledge the European Synchrotron Radiation Facility for beam time allocation, and the staff of beamline BM14 for assistance with data collection. The authors thank A. N. Volkov (Vrije Universiteit Brussel) for revised NMR assignments of cytochrome c and J. Donlon (NUI Galway) and S. S. Wijmenga (Radboud Universiteit Nijmegen) for helpful discussions.

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  1. Nicholas P. Power

    Present address: Present address: Department of Applied Sciences, London South Bank University, London SE1 0AA, UK,

Authors and Affiliations

  1. School of Chemistry, National University of Ireland Galway, University Road, Galway, Ireland

    Róise E. McGovern, Nicholas P. Power & Peter B. Crowley

  2. School of Biochemistry and Immunology, Trinity College, Dublin 2, Ireland

    Humberto Fernandes & Amir R. Khan

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Contributions

P.B.C. conceived and designed the experiments. R.E.M., H.F., A.R.K. and P.B.C. performed the experiments. R.E.M. and P.B.C. analysed the data. N.P.P. contributed the calixarene. P.B.C. wrote the paper. All authors commented on the manuscript.

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Correspondence to Peter B. Crowley.

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McGovern, R., Fernandes, H., Khan, A. et al. Protein camouflage in cytochrome c–calixarene complexes. Nature Chem 4, 527–533 (2012). https://doi.org/10.1038/nchem.1342

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