Vaca L, Kunze D L
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas 77030.
J Membr Biol. 1992 Dec;130(3):241-9. doi: 10.1007/BF00240481.
A large conductance multi-state channel was identified and characterized in single channel recordings from cell-attached and excised patches of the human colonic tumor cell line, T84. The channel activity was dependent on the presence of both permeable cations and anions. In Na(+)-free symmetrical Cl- solutions or Cl(-)-free symmetrical Na+ solutions the channel was inactive. Addition of 5 mM NaCl (NaI or KCl) induced channel activity. The selectivity sequence obtained from the shift in reversal potential was I- (1.9) > Cl- (1) > Na+ (0.5) > K+ (0.3). SO2-(4), SCN- (thiocyanate) and NMDG+ were impermeant. Multiple subconductance states were identified at all voltages explored (+/- 90 mV). The minimum conductance encountered in symmetrical 100 mM NaCl was a 15 pS substrate, the maximum, 210 pS. The channel appeared to be composed of multiples of the 15 pS subunits which were reversibly blocked by the loop diuretic bumetanide (5 microM).
在人结肠肿瘤细胞系T84的细胞贴附式和内翻式膜片的单通道记录中,鉴定并表征了一种大电导多态通道。通道活性依赖于可渗透阳离子和阴离子的同时存在。在无钠对称氯溶液或无氯对称钠溶液中,通道无活性。加入5 mM NaCl(NaI或KCl)可诱导通道活性。从反转电位的偏移获得的选择性顺序为I-(1.9)>Cl-(1)>Na+(0.5)>K+(0.3)。SO42-、SCN-(硫氰酸盐)和NMDG+不可渗透。在所有探索的电压(±90 mV)下都鉴定出了多个亚电导状态。在对称的100 mM NaCl中遇到的最小电导是15 pS的底物,最大电导是210 pS。该通道似乎由15 pS亚基的多个拷贝组成,这些亚基被髓袢利尿剂布美他尼(5 microM)可逆性阻断。