Primary structure, developmental expression and functional properties of an inward rectifier K+ channel of the tunicate.
A cDNA encoding for a tunicate inward rectifier K+ channel (TuIRK) was isolated. TuIRK exhibited the highest similarity (approximately 50%) with mammalian Kir2 (IRK) subfamily. Maternal RNA of TuIRK was detected by RT-PCR in unfertilized eggs. By in situ hybridization, the transcript was observed at the 32-cell stage, restricted at the 64-cell stage in anterior epidermal cells of a4-2 blastomere lineage, and disappeared at the late gastrula stage. Therefore, TuIRK was identified to be the inward rectifier whose expression was previously reported to change dramatically upon the neural/epidermal cell fate selection. In Xenopus oocytes, TuIRK expressed a strongly inward rectifying K+ current. The basic electrophysiological properties of TuIRK were similar to those of the mouse IRK1 (mIRK1), except that the sensitivity to the block by extracellular Mg2+ was much lower than that of mIRK1. To identify the structural determinant, we made mutants of the pore region, and then of the extracellular loop (N226 of TuIRK, and E125 of mIRK1). In E125N mutant of mIRK1, the sensitivity to the Mg2+ block was decreased significantly, whereas N226E of TuIRK1 did not acquire the sensitivity. These results demonstrate the contribution of this site to the Mg2+ block and the presence of additional determinant(s).
Murata
Y
Y
Department of Physiology, Tokyo Medical and Dental University, Graduate School and Faculty of Medicine, D-566, Yushima 1-5-45, Bunkyo, Tokyo 113-8519, Japan.
Okado
H
H
Katsuyama
Y
Y
Okamura
Y
Y
Kubo
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D004553:Electric Conductivity; D004594:Electrophysiology; D015723:Gene Library; D008969:Molecular Sequence Data; D009154:Mutation; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism*; D024661:Potassium Channels, Inwardly Rectifying; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D014561:Urochordata / Q000235:genetics / Q000378:metabolism*
D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying