Tang W, Ruknudin A, Yang W P, Shaw S Y, Knickerbocker A, Kurtz S
Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey 08543-4000, USA.
Mol Biol Cell. 1995 Sep;6(9):1231-40. doi: 10.1091/mbc.6.9.1231.
We describe the expression of gpIRK1, an inwardly rectifying K+ channel obtained from guinea pig cardiac cDNA. gpIRK1 is a homologue of the mouse IRK1 channel identified in macrophage cells. Expression of gpIRK1 in Xenopus oocytes produces inwardly rectifying K+ current, similar to the cardiac inward rectifier current IK1. This current is blocked by external Ba2+ and Cs+. Plasmids containing the gpIRK1 coding region under the transcriptional control of constitutive (PGK) or inducible (GAL) promoters were constructed for expression in Saccharomyces cerevisiae. Several observations suggest that gpIRK1 forms functional ion channels when expressed in yeast. gpIRK1 complements a trk1 delta trk2 delta strain, which is defective in potassium uptake. Expression of gpIRK1 in this mutant restores growth on low potassium media. Growth dependent on gpIRK1 is inhibited by external Cs+. The strain expressing gpIRK1 provides a versatile genetic system for studying the assembly and composition of inwardly rectifying K+ channels.
我们描述了gpIRK1的表达情况,gpIRK1是一种从豚鼠心脏cDNA中获得的内向整流钾通道。gpIRK1是在巨噬细胞中鉴定出的小鼠IRK1通道的同源物。gpIRK1在非洲爪蟾卵母细胞中的表达产生内向整流钾电流,类似于心脏内向整流电流IK1。该电流被细胞外的Ba2+和Cs+阻断。构建了在组成型(PGK)或诱导型(GAL)启动子转录控制下含有gpIRK1编码区的质粒,用于在酿酒酵母中表达。多项观察结果表明,gpIRK1在酵母中表达时形成功能性离子通道。gpIRK1可补充trk1δtrk2δ菌株,该菌株在钾摄取方面存在缺陷。在该突变体中gpIRK1的表达可恢复其在低钾培养基上的生长。依赖gpIRK1的生长受到细胞外Cs+的抑制。表达gpIRK1的菌株为研究内向整流钾通道的组装和组成提供了一个通用的遗传系统。