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Electrophysiological, biochemical, and bioinformatic methods for studying CFTR channel gating and its regulation.

Csanády, L; Vergani, P; Gulyás-Kovács, A; Gadsby, DC; (2011) Electrophysiological, biochemical, and bioinformatic methods for studying CFTR channel gating and its regulation. In: Cystic Fibrosis. (443 - 469). Humana Press: New York, US. Green open access

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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.

Type: Book chapter
Title: Electrophysiological, biochemical, and bioinformatic methods for studying CFTR channel gating and its regulation.
Location: US
ISBN: 978-1-61779-116-1
Open access status: An open access version is available from UCL Discovery
DOI: 10.1007/978-1-61779-117-8_28
Publisher version: http://dx.doi.org/10.1007/978-1-61779-117-8_28
Language: English
Additional information: © Springer Science+Business Media, LLC 2011 The final publication is available at http://link.springer.com/protocol/10.1007/978-1-61779-117-8_28
Keywords: Animals, Biochemistry, Cell Membrane, Computational Biology, Cysteine, Cystic Fibrosis Transmembrane Conductance Regulator, Electric Conductivity, Electrophysiological Processes, Evolution, Molecular, Humans, Ion Channel Gating, Kinetics, Microelectrodes, Mutation, Oocytes, Patch-Clamp Techniques, Phosphorylation, Plasmids, Protein Structure, Tertiary, Reproducibility of Results, Temperature, Thermodynamics, Xenopus
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Neuro, Physiology and Pharmacology
URI: https://discovery.ucl.ac.uk/id/eprint/1307803
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