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Probing CFTR Channel Structure and Function Using the Substituted-Cysteine-Accessibility Method

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Cystic Fibrosis Methods and Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 70))

Abstract

The cystic fibrosis transmembrane conductance regulator (CFTR) forms a chloride channel whose activation is regulated by phosphorylation and by ATP binding and hydrolysis (1-4). The functional properties of the channel have been extensively studied using electrophysiological techniques (4). Less is known about the structural bases for the functional properties. The cloning of CFTR in 1989 provided the primary amino acid sequence and a putative transmembrane topology of the protein (5). In order to understand the structural bases for the functional properties of the channel, we sought to identify the residues lining the ion channel because they are likely to be the major determinants of the channel’s functional properties. Although the channel-lining residues lie within membrane-spanning segments, they are part of the water-accessible surface of the protein. The substituted-cysteine-accessibility method (SCAM), which we developed, provides an approach to identify systematically the channel-lining residues (6-9). We have applied SCAM to three of CFTR’s 12 putative membrane-spanning segments (9-11).

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Author information

Authors and Affiliations

  1. Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, NY

    Myles H. Akabas

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  1. Myles H. Akabas
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Editor information

Editors and Affiliations

  1. Division of Molecular Medicine, Oregon Health Sciences University, Portland, OR

    William R. Skach MD

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© 2002 Humana Press Inc.

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Akabas, M.H. (2002). Probing CFTR Channel Structure and Function Using the Substituted-Cysteine-Accessibility Method. In: Skach, W.R. (eds) Cystic Fibrosis Methods and Protocols. Methods in Molecular Medicine™, vol 70. Humana Press. https://doi.org/10.1385/1-59259-187-6:159

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  • DOI: https://doi.org/10.1385/1-59259-187-6:159

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-897-4

  • Online ISBN: 978-1-59259-187-9

  • eBook Packages: Springer Protocols

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