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Cysteine-Based Cross-Linking Approach to Study Inter-domain Interactions in Ion Channels

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Ion Channels

Part of the book series: Methods in Molecular Biology ((MIMB,volume 998))

Abstract

Cysteine contains a highly reactive thiol group and therefore under oxidizing conditions a disulfide bond can form between a pair of cysteines that are juxtaposed in the close vicinity, which can be only reversed by reducing agents. These attributes have been elegantly exploited to study the functional role of an interaction or contact between two adjacent domains that are present in ion channels or virtually in any proteins, by introducing double cysteine substitutions at the domain interface and measuring changes in the ion channel functions arising from cross-linking the two substituted cysteines via formation of a disulfide bond. Here I describe this cysteine-based cross-linking approach.

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Author information

Authors and Affiliations

  1. Faculty of Biological Sciences, School of Biomedical Sciences, University of Leeds, Leeds, UK

    Lin-Hua Jiang

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  1. Lin-Hua Jiang
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Editor information

Editors and Affiliations

  1. School of Biomedical Sciences, University of Leeds, Garstang Bld. 6.31d, Leeds, LS2 9JT, United Kingdom

    Nikita Gamper

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Jiang, LH. (2013). Cysteine-Based Cross-Linking Approach to Study Inter-domain Interactions in Ion Channels. In: Gamper, N. (eds) Ion Channels. Methods in Molecular Biology, vol 998. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-351-0_21

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  • DOI: https://doi.org/10.1007/978-1-62703-351-0_21

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-350-3

  • Online ISBN: 978-1-62703-351-0

  • eBook Packages: Springer Protocols

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