Department of Physics and Biophysics, Wrocław University of Environmental and Life Sciences, Norwida 25, PL-50-375 Wrocław, Poland.
C R Biol. 2012 Dec;335(12):722-30. doi: 10.1016/j.crvi.2012.11.004. Epub 2012 Dec 21.
The patch-clamp technique was used to examine the effect of trimethyllead chloride (Met(3)PbCl) on SV channel activity in red beet (Beta vulgaris L.) taproot vacuoles. It was found that in the control bath the macroscopic currents showed the typical slow activation and a strong outward rectification of the steady-state currents. An addition of Met(3)PbCl to the bath solution blocked, in a concentration-dependent manner, SV currents in red beet vacuoles. The time constant τ increased several times in the presence of 100 μM trimethyllead chloride at all voltages tested. When single channel properties were analyzed, only little channel activity could be recorded in the presence of 100 μM Met(3)PbCl. Trimethyllead chloride decreased significantly (by about one order of magnitude) the open probability of single channels. The recordings of single channel activity obtained in the presence and absence of Met(3)PbCl showed that organolead only slightly (by ca. 10%) decreased the unitary conductance of single channels. It was also found that Met(3)PbCl diminished significantly the number of SV channel openings, whereas it did not change the opening times of the channels. Taken together, these results suggest that Met(3)PbCl binding site is located outside the channel selectivity filter.
采用膜片钳技术研究了三甲基氯化铅(Met(3)PbCl)对甜菜(Beta vulgaris L.)主根液泡中 SV 通道活性的影响。结果发现,在对照浴液中,宏观电流表现出典型的缓慢激活和强外向整流的稳态电流。在 100 μM 三甲基氯化铅存在的情况下,添加到浴液中的 Met(3)PbCl 以浓度依赖的方式阻断了甜菜液泡中的 SV 电流。在所有测试的电压下,100 μM 三甲基氯化铅存在时,时间常数 τ 增加了几倍。当分析单通道特性时,在 100 μM Met(3)PbCl 的存在下,只能记录到很少的通道活性。三甲基氯化铅显著降低了单通道的开放概率(约降低一个数量级)。在有和没有 Met(3)PbCl 存在的情况下获得的单通道活动记录表明,有机铅仅略微(约 10%)降低了单通道的单位电导。还发现 Met(3)PbCl 显著减少了 SV 通道开放的数量,而不改变通道的开放时间。总之,这些结果表明,Met(3)PbCl 结合位点位于通道选择性滤器之外。