Makhina E N, Kelly A J, Lopatin A N, Mercer R W, Nichols C G
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110.
J Biol Chem. 1994 Aug 12;269(32):20468-74.
A complementary DNA encoding an inward rectifier K+ channel (HRK1) was isolated from human hippocampus using a 392-base pair cDNA (HHCMD37) as a probe. HRK1 shows sequence similarity to three recently cloned inwardly rectifying potassium channels (IRK1, GIRK1, and ROMK1, 60, 42, and 37%, respectively) and has a similar proposed topology of two membrane spanning domains that correspond to the inner core structure of voltage gated K+ channels. When HRK1 was expressed in Xenopus oocytes, large inward K+ currents were observed below the K+ reversal potential but very little outward K+ current was observed. In on-cell membrane patches, single channel conductance (g) was estimated to be 10 picosiemens by both direct measurement and noise analysis, in 102 mM external [K+]. HRK1 currents were blocked by external Ba2+ and Cs+ (K(0) = 183 microM, and K(-130) = 30 microM, respectively), and internal tetraethylammonium ion (K(0) = 62 microM), but were insensitive to external tetraethylammonium ion. The functional properties of HRK1 are very similar to those of glial cell inward rectifier K+ channels and HRK1 may represent a glial cell inward rectifier.
利用一段392个碱基对的互补DNA(HHCMD37)作为探针,从人脑海马体中分离出一种编码内向整流钾通道(HRK1)的互补DNA。HRK1与最近克隆的三种内向整流钾通道(IRK1、GIRK1和ROMK1,序列相似性分别为60%、42%和37%)具有序列相似性,并且具有类似的推测拓扑结构,即两个跨膜结构域,这与电压门控钾通道的内核结构相对应。当HRK1在非洲爪蟾卵母细胞中表达时,在钾离子反转电位以下观察到较大的内向钾电流,但向外的钾电流很少。在细胞膜片上,通过直接测量和噪声分析,在102 mM的外部[K+]条件下,单通道电导(g)估计为10皮西门子。HRK1电流被外部的Ba2+和Cs+(K(0)分别为183 microM和K(-130)为30 microM)以及内部的四乙铵离子(K(0)为62 microM)阻断,但对外部四乙铵离子不敏感。HRK1的功能特性与胶质细胞内向整流钾通道非常相似,HRK1可能代表一种胶质细胞内向整流器。