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Science 23 June 1989:
Vol. 244. no. 4911, pp. 1469 - 1472
DOI: 10.1126/science.2472007
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Articles

Science, Vol 244, Issue 4911, 1469-1472
Copyright © 1989 by American Association for the Advancement of Science

articles

Purification and reconstitution of chloride channels from kidney and trachea

DW Landry, MH Akabas, C Redhead, A Edelman, EJ Cragoe Jr, and Q Al-Awqati

Department of Medicine, College of Physicians and Surgeons, Columbia University, New York, NY 10032.

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. Indanyl-oxyacetic 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.


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Science. ISSN 0036-8075 (print), 1095-9203 (online)