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脑线粒体中 ATP 调节钾通道的单通道研究。

Single channel studies of the ATP-regulated potassium channel in brain mitochondria.

机构信息

Department of Biophysics, Warsaw University of Life Sciences SGGW, 159 Nowoursynowska St., 02-776, Warsaw, Poland.

出版信息

J Bioenerg Biomembr. 2009 Aug;41(4):323-34. doi: 10.1007/s10863-009-9233-7. Epub 2009 Oct 10.

Abstract

Mitochondrial potassium channels in the brain have been suggested to have an important role in neuroprotection. The single channel activity of mitochondrial potassium channels was measured after reconstitution of the purified inner membrane from rat brain mitochondria into a planar lipid bilayer. In addition to a large conductance potassium channel that was described previously, we identified a potassium channel that has a mean conductance of 219 +/- 15 pS. The activity of this channel was inhibited by ATP/Mg(2+) and activated by the potassium channel opener BMS191095. Channel activity was not influenced either by 5-hydroxydecanoic acid, an inhibitor of mitochondrial ATP-regulated potassium channels, or by the plasma membrane ATP-regulated potassium channel blocker HMR1098. Likewise, this mitochondrial potassium channel was unaffected by the large conductance potassium channel inhibitor iberiotoxin or by the voltage-dependent potassium channel inhibitor margatoxin. The amplitude of the conductance was lowered by magnesium ions, but the opening ability was unaffected. Immunological studies identified the Kir6.1 channel subunit in the inner membrane from rat brain mitochondria. Taken together, our results demonstrate for the first time the single channel activity and properties of an ATP-regulated potassium channel from rat brain mitochondria.

摘要

脑线粒体钾通道被认为在神经保护中具有重要作用。从大鼠脑线粒体纯化的内膜重新构成平面脂双层后,测量了线粒体钾通道的单通道活性。除了先前描述的大电导钾通道外,我们还鉴定出一种钾通道,其平均电导为 219±15pS。该通道的活性受 ATP/Mg(2+)抑制,并被钾通道开放剂 BMS191095 激活。5-羟癸酸(一种线粒体 ATP 调节性钾通道抑制剂)或质膜 ATP 调节性钾通道阻滞剂 HMR1098 对通道活性均无影响。同样,这种线粒体钾通道不受大电导钾通道抑制剂 Iberiotoxin 或电压依赖性钾通道抑制剂 Margatoxin 的影响。镁离子降低电导幅度,但不影响通道的开启能力。免疫研究在大鼠脑线粒体的内膜中鉴定出 Kir6.1 通道亚基。综上所述,我们的研究结果首次证明了大鼠脑线粒体中存在一种 ATP 调节性钾通道的单通道活性和特性。

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