Purification of a Cl-(-)channel protein of sarcoplasmic reticulum by assaying the channel activity in the planar lipid bilayer system.
A Cl- channel protein of sarcoplasmic reticulum (SR) was purified by assaying the channel activity in a planar lipid bilayer system. The light fraction of SR vesicles was solubilized in CHAPS and fractioned by anion exchange, gel filtration, and affinity chromatography with concanavalin A. All fractions in each step were reconstituted into vesicles with asolectin by dialysis and their channel activities were assayed after the vesicles had been fused into a planar lipid bilayer. A 100-kDa protein, different from Ca2(+)-ATPase, was found to form anion channels.
Ide
T
T
Department of Biophysical Engineering, Faculty of Engineering Science, Osaka University, Japan.
Sakamoto
H
H
Morita
T
T
Taguchi
T
T
Kasai
M
M
D000818:Animals; D018118:Chloride Channels; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004553:Electric Conductivity; D004591:Electrophoresis, Polyacrylamide Gel; D007473:Ion Channels / Q000502:physiology; D008051:Lipid Bilayers; D008565:Membrane Proteins / Q000187:drug effects / Q000302:isolation & purification / Q000502:physiology*; D008970:Molecular Weight; D009132:Muscles / Q000502:physiology; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000502:physiology*; D013431:Sulfates / Q000494:pharmacology
D018118:Chloride Channels; D007473:Ion Channels; D008051:Lipid Bilayers; D008565:Membrane Proteins; D013431:Sulfates