New mammalian chloride channel identified by expression cloning.
Ion channels selectively permeable to chloride ions regulate cell functions as diverse as excitability and control of cell volume. Using expression cloning techniques, a complementary DNA from an epithelial cell line has been isolated, sequenced and its putative structure examined by site-directed mutagenesis. This cDNA, encoding a 235-amino-acid protein, gave rise to a chloride-selective outward current when expressed in Xenopus oocytes. The expressed, outwardly rectifying chloride current was calcium-insensitive and was blocked by nucleotides applied to the cell surface. Mutation of a putative nucleotide-binding site resulted in loss of nucleotide block but incurred dependence on extracellular calcium concentration. The unusual sequence of this putative channel protein suggests a new class of ion channels not related to other previously cloned chloride channels.
Paulmichl
M
M
Department of Pharmacology, Mayo Foundation, Rochester, Minnesota 55905.
Li
Y
Y
Wickman
K
K
Ackerman
M
M
Peralta
E
E
Clapham
D
D
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D018118:Chloride Channels; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004285:Dogs; D004553:Electric Conductivity; D015870:Gene Expression; D007668:Kidney; D007700:Kinetics; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009865:Oocytes / Q000378:metabolism; D010957:Plasmids; D014162:Transfection; D014982:Xenopus laevis
D018118:Chloride Channels; D008565:Membrane Proteins; D004247:DNA