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Optochemical control of genetically engineered neuronal nicotinic acetylcholine receptors

Nature Chemistry volume 4, pages 105–111 (2012)Cite this article

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Abstract

Advances in synthetic chemistry, structural biology, molecular modelling and molecular cloning have enabled the systematic functional manipulation of transmembrane proteins. By combining genetically manipulated proteins with light-sensitive ligands, innately ‘blind’ neurobiological receptors can be converted into photoreceptors, which allows them to be photoregulated with high spatiotemporal precision. Here, we present the optochemical control of neuronal nicotinic acetylcholine receptors (nAChRs) with photoswitchable tethered agonists and antagonists. Using structure-based design, we produced heteromeric α3β4 and α4β2 nAChRs that can be activated or inhibited with deep-violet light, but respond normally to acetylcholine in the dark. The generation of these engineered receptors should facilitate investigation of the physiological and pathological functions of neuronal nAChRs and open a general pathway to photosensitizing pentameric ligand-gated ion channels.

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Figure 1: Photoswitchable ligand design for the optical control of neuronal nAChRs.
Figure 2: Choice of cysteine attachment sites for photoswitchable ligand conjugation.
Figure 3: Photoactivation of nAChRs with a tethered agonist.
Figure 4: Molecular model of a nAChR with a virtually tethered photoswitchable agonist.
Figure 5: Photoinhibition of nAChRs with a tethered antagonist.
Figure 6: Thermal relaxation of the photoswitchable agonist and antagonist.

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Acknowledgements

Support for the work was provided by the Nanomedicine Development Center for the Optical Control of Biological Function PN2EY018241 (D.T., R.H.K.), The European Research Commission Advanced Grant (D.T.) and the Deutsche Forschungsgemeinschaft SFB 749 (D.T.).

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Author notes

  1. Ivan Tochitsky and Matthew R. Banghart: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Molecular and Cell Biology, University of California, Berkeley, 94720, California, USA

    Ivan Tochitsky, Alexandre Mourot & Richard H. Kramer

  2. Department of Chemistry, University of California, Berkeley, 94720, California, USA

    Matthew R. Banghart, Jennifer Z. Yao & Dirk Trauner

  3. Helen Wills Neuroscience Institute, University of California, Berkeley, 94720, California, USA

    Benjamin Gaub & Richard H. Kramer

  4. Department of Chemistry, Ludwig-Maximilians-Universität, München and Center for Integrated Protein Science, Munich, 81377, Germany

    Dirk Trauner

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Contributions

M.R.B., A.M., R.H.K. and D.T. designed the research. M.R.B. and J.Z.Y. synthesized the ligands. I.T., A.M. and B.G. performed the research and analysed the data. I.T., M.R.B. and D.T. co-wrote the paper.

Corresponding authors

Correspondence to Richard H. Kramer or Dirk Trauner.

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The authors declare no competing financial interests.

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Tochitsky, I., Banghart, M., Mourot, A. et al. Optochemical control of genetically engineered neuronal nicotinic acetylcholine receptors. Nature Chem 4, 105–111 (2012). https://doi.org/10.1038/nchem.1234

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