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Molecular tweezers modulate 14-3-3 protein–protein interactions

Nature Chemistry volume 5, pages 234–239 (2013)Cite this article

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Abstract

Supramolecular chemistry has recently emerged as a promising way to modulate protein functions, but devising molecules that will interact with a protein in the desired manner is difficult as many competing interactions exist in a biological environment (with solvents, salts or different sites for the target biomolecule). We now show that lysine-specific molecular tweezers bind to a 14-3-3 adapter protein and modulate its interaction with partner proteins. The tweezers inhibit binding between the 14-3-3 protein and two partner proteins—a phosphorylated (C-Raf) protein and an unphosphorylated one (ExoS)—in a concentration-dependent manner. Protein crystallography shows that this effect arises from the binding of the tweezers to a single surface-exposed lysine (Lys214) of the 14-3-3 protein in the proximity of its central channel, which normally binds the partner proteins. A combination of structural analysis and computer simulations provides rules for the tweezers' binding preferences, thus allowing us to predict their influence on this type of protein–protein interactions.

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Figure 1: Molecular tweezers.
Figure 2: Molecular tweezers binding to 14-3-3.
Figure 3: Competition between molecular tweezers and FAM-C-RafpS259 (green) or FAM-ExoS (black) with respect to their binding sites on 14-3-3σ.
Figure 4: Structure of molecular tweezers bound to 14-3-3σ.
Figure 5: Prerequisites for efficient surface lysine complexation by molecular tweezers in proteins.
Figure 6: Relative position of the molecular tweezers' binding site compared to C-RafpS259 and ExoS.

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Acknowledgements

C.O. thanks AstraZeneca, Bayer CropScience, Bayer Healthcare, Boehringer Ingelheim and Merck KGaA for support. E.S-G acknowledges a Liebig stipend and K.B-R. a pre-doctoral stipend from the Fonds der Chemischen Industrie. E.S-G thanks the Cluster of Excellence RESOLV (EXC 1069) funded by the Deutsche Forschungsgemeinschaft for financial support and W. Thiel for useful suggestions. The authors thank E. Hofmann, F. Syberg, I. Vetter and the SLS beamline staff for data collection at the Swiss Light Source, beamline PXII-X10SA, Paul Scherrer Institute, Villigen, Switzerland.

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  1. David Bier and Rolf Rose: These authors contributed equally to this work

Authors and Affiliations

  1. Chemical Genomics Centre of the Max-Planck-Society, Otto-Hahn-Strasse 15, 44227, Dortmund, Germany

    David Bier, Rolf Rose, Maria Bartel & Christian Ottmann

  2. Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470, Mülheim an der Ruhr, Germany

    Kenny Bravo-Rodriguez, Juan Manuel Ramirez-Anguita & Elsa Sanchez-Garcia

  3. Department of Chemistry, University of Duisburg-Essen, Universitätsstrasse 7, 45117, Essen, Germany

    Som Dutt, Constanze Wilch, Frank-Gerrit Klärner & Thomas Schrader

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Contributions

D.B., R.R. and M.B. carried out the experiments. K.B.-R. and J.M.R.-A. performed the QM/MM and MD calculations. S.D. and C.W. synthesized the tweezers. F.-G.K. designed their synthesis. C.O., T.S. and E.S.-G. designed the experiments and calculations and wrote the paper.

Corresponding authors

Correspondence to Elsa Sanchez-Garcia, Thomas Schrader or Christian Ottmann.

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Bier, D., Rose, R., Bravo-Rodriguez, K. et al. Molecular tweezers modulate 14-3-3 protein–protein interactions. Nature Chem 5, 234–239 (2013). https://doi.org/10.1038/nchem.1570

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