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Cystic Fibrosis pp 443–469Cite as

Electrophysiological, Biochemical, and Bioinformatic Methods for Studying CFTR Channel Gating and Its Regulation

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 741))

Abstract

CFTR is the only member of the ABC (ATP-binding cassette) protein superfamily known to function as an ion channel. Most other ABC proteins are ATP-driven transporters, in which a cycle of ATP binding and hydrolysis, at intracellular nucleotide binding domains (NBDs), powers uphill substrate translocation across the membrane. In CFTR, this same ATP-driven cycle opens and closes a transmembrane pore through which chloride ions flow rapidly down their electrochemical gradient. Detailed analysis of the pattern of gating of CFTR channels thus offers the opportunity to learn about mechanisms of function not only of CFTR channels but also of their ABC transporter ancestors. In addition, CFTR channel gating is subject to complex regulation by kinase-mediated phosphorylation at multiple consensus sites in a cytoplasmic regulatory domain that is unique to CFTR. Here we offer a practical guide to extract useful information about the mechanisms that control opening and closing of CFTR channels: on how to plan (including information obtained from analysis of multiple sequence alignments), carry out, and analyze electrophysiological and biochemical experiments, as well as on how to circumvent potential pitfalls.

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Notes

  1. 1.

    After reducing the cyclic gating scheme to a closed-to-open equilibrium by impairing hydrolysis, this equilibrium constant can be obtained from P o, as K eq= P o/(1–P o)

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Author information

Authors and Affiliations

  1. Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary

    László Csanády

  2. Department of Neuroscience, Physiology and Pharmacology, University College London, London, UK

    Paola Vergani

  3. Laboratory of Cardiac/Membrane Physiology, The Rockefeller University, New York, 10065, NY, USA

    Attila Gulyás-Kovács

  4. Laboratory of Cardiac/Membrane Physiology, The Rockefeller University, 10065, New York, NY, USA

    David C. Gadsby

Authors
  1. László Csanády
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  2. Paola Vergani
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  3. Attila Gulyás-Kovács
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  4. David C. Gadsby
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Corresponding authors

Correspondence to László Csanády or David C. Gadsby .

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Editors and Affiliations

  1. Faculty of Sciences, Centre for Biodiversity & Functional, University of Lisboa, Campo Grande, Lisboa, 1749-016, Portugal

    Margarida D. Amaral

  2. , Department of Physiology, University of Regensburg, Universitätsstr. 31, Regensburg, 93053, Germany

    Karl Kunzelmann

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Csanády, L., Vergani, P., Gulyás-Kovács, A., Gadsby, D.C. (2011). Electrophysiological, Biochemical, and Bioinformatic Methods for Studying CFTR Channel Gating and Its Regulation. In: Amaral, M., Kunzelmann, K. (eds) Cystic Fibrosis. Methods in Molecular Biology, vol 741. Humana Press. https://doi.org/10.1007/978-1-61779-117-8_28

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  • DOI: https://doi.org/10.1007/978-1-61779-117-8_28

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