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Generation of functional ion-channel tools by E3 targeting

Nature Biotechnology volume 23, pages 1289–1293 (2005)Cite this article

Abstract

Here we describe a strategy for generating ion-channel inhibitors. It takes advantage of antibody specificity combined with a pattern recognition approach that targets the third extracellular region (E3) of a channel. To test the concept, we first focused on TRPC5, a member of the transient receptor potential (TRP) calcium channel family, the study of which has been hindered by poor pharmacological tools. Extracellular application of E3-targeted anti-TRPC5 antibody led to a specific TRPC5 inhibitor, enabling TRPC5 to be distinguished from its closest family members, and TRPC5 function to be explored in a relatively intractable physiological system. E3 targeting was further applied to voltage-gated sodium channels, leading to discovery of a subtype-specific inhibitor of NaV1.5. These examples illustrate the potential power of E3 targeting as a systematic method for producing gene-type specific ion-channel inhibitors for use in routine assays on cells or tissues from a range of species and having therapeutic potential.

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Figure 1: E3 targeting.
Figure 2: TRPC5 inhibitor.
Figure 3: Physiological application.
Figure 4: Generalization to sodium channels.

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Acknowledgements

We are grateful to C. Montell, J. Putney, Y. Mori, A. Srivastava and T. Zimmer for TRPC1, TRPC4, TRPC5, NaV1.4/5 cDNAs, and A.N. Bateson and J. Colyer for critical reading of the manuscript. The work was supported by the Wellcome Trust and British Heart Foundation.

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Authors and Affiliations

  1. Membrane and Systems Biology Research Institute, University of Leeds, Leeds, LS2 9JT, UK

    Shang-Zhong Xu, Fanning Zeng, Jing Li, Asipu Sivaprasadarao & David J Beech

  2. University Laboratory of Physiology, University of Oxford, OX1 3PT, UK

    Ming Lei

  3. Rheumatology Unit, Institute of Child Health, University College London, London, WC1N 1EH, UK

    Bin Gao

  4. Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, P.R. China

    Chenliang Xiong

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Correspondence to David J Beech.

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Xu, SZ., Zeng, F., Lei, M. et al. Generation of functional ion-channel tools by E3 targeting. Nat Biotechnol 23, 1289–1293 (2005). https://doi.org/10.1038/nbt1148

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