Purification and reconstitution of chloride channels from kidney and trachea

Donald W. Landry, Myles H. Akabas, Christopher Redhead, Aleksander Edelman, Edward J. Cragoe, Qais Al-Awqati

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Abstract

Chloride channels mediate absorption and secretion of fluid in epithelia, and the regulation of these channels is now known to be defective in cystic fibrosis. Indanyloxyacetic acid 94 (IAA-94) is a high-affinity ligand for the chloride channel, and an affinity resin based on that structure was developed. Solubilized proteins from kidney and trachea membranes were applied to the affinity matrix, and four proteins with apparent molecular masses of 97, 64, 40, and 27 kilodaltons were eluted from the column by excess IAA-94. A potential-dependent 36Cl- uptake was observed after reconstituting these proteins into liposomes. Three types of chloride channels with single-channel conductances of 26, 100, and 400 picosiemens were observed after fusion of these liposomes with planar lipid bilayers. Similar types of chloride channels have been observed in epithelia.

Original languageEnglish (US)
Pages (from-to)1469-1472
Number of pages4
JournalScience
Volume244
Issue number4911
DOIs
StatePublished - Jan 1 1989
Externally publishedYes

ASJC Scopus subject areas

  • General

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    Landry, D. W., Akabas, M. H., Redhead, C., Edelman, A., Cragoe, E. J., & Al-Awqati, Q. (1989). Purification and reconstitution of chloride channels from kidney and trachea. Science, 244(4911), 1469-1472. https://doi.org/10.1126/science.2472007