Purification and reconstitution of chloride channels from kidney and trachea.
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.
Landry
D W
DW
Department of Medicine, College of Physicians and Surgeons, Columbia University, New York, NY 10032.
Akabas
M H
MH
Redhead
C
C
Edelman
A
A
Cragoe
E J
EJ
Al-Awqati
Q
Q
D000818:Animals; D001436:Bacteriorhodopsins / Q000378:metabolism; D002417:Cattle; D018118:Chloride Channels; D002712:Chlorides / Q000302:isolation & purification* / Q000502:physiology; D002713:Chlorine / Q000378:metabolism; D002846:Chromatography, Affinity; D004553:Electric Conductivity; D004591:Electrophoresis, Polyacrylamide Gel; D007189:Indans; D007473:Ion Channels / Q000502:physiology; D007672:Kidney Cortex / Q000032:analysis*; D008027:Light; D008081:Liposomes / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000502:physiology; D008970:Molecular Weight; D011868:Radioisotopes; D014132:Trachea / Q000032:analysis*; D014634:Valinomycin / Q000494:pharmacology
D018118:Chloride Channels; D002712:Chlorides; D007189:Indans; D007473:Ion Channels; D008081:Liposomes; D008565:Membrane Proteins; D011868:Radioisotopes; C060056:indanoxyacetic acid 94; D014634:Valinomycin; D002713:Chlorine; D001436:Bacteriorhodopsins