Duke Institute of Molecular Physiology, Duke University Medical Center, Durham, North Carolina 27704, USA.
J Biol Chem. 2013 Aug 9;288(32):23128-40. doi: 10.1074/jbc.M113.491654. Epub 2013 Jun 20.
Recent studies have shown that the pyruvate-isocitrate cycling pathway, involving the mitochondrial citrate/isocitrate carrier and the cytosolic NADP-dependent isocitrate dehydrogenase (ICDc), is involved in control of glucose-stimulated insulin secretion (GSIS). Here we demonstrate that pyruvate-isocitrate cycling regulates expression of the voltage-gated potassium channel family member Kv2.2 in islet β-cells. siRNA-mediated suppression of ICDc, citrate/isocitrate carrier, or Kv2.2 expression impaired GSIS, and the effect of ICDc knockdown was rescued by re-expression of Kv2.2. Moreover, chronic exposure of β-cells to elevated fatty acids, which impairs GSIS, resulted in decreased expression of Kv2.2. Surprisingly, knockdown of ICDc or Kv2.2 increased rather than decreased outward K(+) current in the 832/13 β-cell line. Immunoprecipitation studies demonstrated interaction of Kv2.1 and Kv2.2, and co-overexpression of the two channels reduced outward K(+) current compared with overexpression of Kv2.1 alone. Also, siRNA-mediated knockdown of ICDc enhanced the suppressive effect of the Kv2.1-selective inhibitor stromatoxin1 on K(+) currents. Our data support a model in which a key function of the pyruvate-isocitrate cycle is to maintain levels of Kv2.2 expression sufficient to allow it to serve as a negative regulator of Kv channel activity.
最近的研究表明,涉及线粒体柠檬酸/异柠檬酸载体和细胞质 NADP 依赖性异柠檬酸脱氢酶(ICDc)的丙酮酸-异柠檬酸循环途径参与了葡萄糖刺激的胰岛素分泌(GSIS)的控制。在这里,我们证明了丙酮酸-异柠檬酸循环调节胰岛β细胞中电压门控钾通道家族成员 Kv2.2 的表达。siRNA 介导的 ICDc、柠檬酸/异柠檬酸载体或 Kv2.2 表达抑制损害了 GSIS,而 ICDc 敲低的作用可以通过重新表达 Kv2.2 来挽救。此外,β细胞长期暴露于升高的脂肪酸会损害 GSIS,导致 Kv2.2 的表达减少。令人惊讶的是,ICDc 或 Kv2.2 的敲低反而增加了 832/13 β 细胞系中的外向 K(+)电流,而不是减少了外向 K(+)电流。免疫沉淀研究表明 Kv2.1 和 Kv2.2 之间存在相互作用,并且这两个通道的共表达与单独过表达 Kv2.1 相比,减少了外向 K(+)电流。此外,siRNA 介导的 ICDc 敲低增强了 Kv2.1 选择性抑制剂 stromatoxin1 对 K(+)电流的抑制作用。我们的数据支持这样一种模型,即丙酮酸-异柠檬酸循环的一个关键功能是维持 Kv2.2 表达水平,使其足以作为 Kv 通道活性的负调节剂。