Halide permeation through three types of epithelial anion channels after reconstitution into giant liposomes.
Anion-selective channels from apical membranes of cultured CFPAC-1 cells were isolated and incorporated into giant liposomes for patch clamp recording. Liposomes were formed from L-alpha-lecithin by a dehydration-hydration method. Ion channels were characterized using the excised inside-out patch clamp configuration. The most commonly observed anion channels were similar to those observed in native epithelial tissues. The linear 20 pS Cl- channel had the halide permeability sequence Cl- > I- > or = Br- > F-, and showed anomalous mole-fraction behavior in solutions containing different proportions of Cl- and F- ions. The autwardly rectifying Cl- channel had the halide permeability sequence I- > Br- > Cl- > F-, and also showed anomalous mole-fraction behavior, indicating that both these channels probably contain multi-ion pores. The third, voltage-dependent anion channel showed at least five different substrates, had a conductance of 390 pS in the main state, and showed two types of kinetics, fast (openings and closings < 1 ms), and slow (openings and closings > 1 s). The channel was seen more frequently after reconstitution into giant liposomes than in intact cells. It was not selective amongst the halides, and there was no deviation from a linear dependence of relative current on molar fractions, indicating relatively simple permeation through the pore. Differences in halide permeabilities suggest that different anion channels may be related to different membrane proteins. Comparison with the chloride channel proteins isolated biochemically from epithelial cell membranes is discussed.
Duszyk
M
M
Department of Medicine, University of Alberta, Edmonton, Canada.
Liu
D
D
French
A S
AS
Man
S F
SF
D002463:Cell Membrane Permeability / Q000502:physiology; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D018118:Chloride Channels; D004848:Epithelium / Q000378:metabolism; D006219:Halogens / Q000493:pharmacokinetics*; D006801:Humans; D007473:Ion Channels / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D008081:Liposomes; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D010190:Pancreatic Neoplasms / Q000378:metabolism; D014407:Tumor Cells, Cultured
D018118:Chloride Channels; D006219:Halogens; D007473:Ion Channels; D008081:Liposomes; D008565:Membrane Proteins