Koszela-Piotrowska Izabela, Matkovic Karolina, Szewczyk Adam, Jarmuszkiewicz Wieslawa
Laboratory of Intracellular Ion Channels, Nencki Institute of Experimental Biology, 3 Pasteur st., 02-093 Warsaw, Poland.
Biochem J. 2009 Nov 11;424(2):307-16. doi: 10.1042/BJ20090991.
In the present study, we describe the existence of a novel potassium channel in the plant [potato (Solanum tuberosum) tuber] mitochondrial inner membrane. We found that substances known to modulate large-conductance calcium-activated potassium channel activity influenced the bioenergetics of potato tuber mitochondria. In isolated mitochondria, Ca2+ and NS1619 {1,3-dihydro-1-[2-hydroxy-5-(trifluoromethyl)phenyl]-5-(trifluoromethyl)-2H-ben-zimidazole-2-one; a potassium channel opener} were found to depolarize the mitochondrial membrane potential and to stimulate resting respiration. These effects were blocked by iberiotoxin (a potassium channel inhibitor) in a potassium-dependent manner. Additionally, the electrophysiological properties of the large-conductance potassium channel present in the potato tuber inner mitochondrial membrane are described in a reconstituted system, using planar lipid bilayers. After incorporation in 50/450 mM KCl gradient solutions, we recorded large-conductance potassium channel activity with conductance from 502+/-15 to 615+/-12 pS. The probability of channel opening was increased by Ca2+ and reduced by iberiotoxin. Immunological analysis with antibodies raised against the mammalian plasma-membrane large-conductance Ca2+-dependent K+ channel identified a pore-forming alpha subunit and an auxiliary beta2 subunit of the channel in potato tuber mitochondrial inner membrane. These results suggest that a large-conductance calcium-activated potassium channel similar to that of mammalian mitochondria is present in potato tuber mitochondria.
在本研究中,我们描述了植物[马铃薯(Solanum tuberosum)块茎]线粒体内膜中一种新型钾通道的存在。我们发现,已知可调节大电导钙激活钾通道活性的物质会影响马铃薯块茎线粒体的生物能量学。在分离的线粒体中,发现Ca2+和NS1619 {1,3-二氢-1-[2-羟基-5-(三氟甲基)苯基]-5-(三氟甲基)-2H-苯并咪唑-2-酮;一种钾通道开放剂}会使线粒体膜电位去极化并刺激静息呼吸。这些效应以钾依赖的方式被iberiotoxin(一种钾通道抑制剂)阻断。此外,使用平面脂质双层在重构系统中描述了马铃薯块茎线粒体内膜中存在的大电导钾通道的电生理特性。在加入50/450 mM KCl梯度溶液后,我们记录到了电导为502±15至615±12 pS的大电导钾通道活性。通道开放的概率因Ca2+而增加,因iberiotoxin而降低。用针对哺乳动物质膜大电导钙依赖性钾通道产生的抗体进行免疫分析,鉴定出马铃薯块茎线粒体内膜中该通道的一个形成孔道的α亚基和一个辅助β2亚基。这些结果表明,马铃薯块茎线粒体中存在一种与哺乳动物线粒体类似的大电导钙激活钾通道。