Changes in the Mitochondrial Proteome during the Anoxia to Air Transition in Rice Focus around Cytochrome-containing Respiratory Complexes*
Changes in the Mitochondrial Proteome during the Anoxia to Air Transition in Rice Focus around Cytochrome-containing Respiratory Complexes*
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DOI：
10.1074/jbc.m406015200
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发表时间：
2004-09
影响因子：
4.8
通讯作者：
Harvey Millar;Alice E Trend;J. Heazlewood
中科院分区：
文献类型：
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作者：
Harvey Millar;Alice E Trend;J. Heazlewood
水稻幼苗在缺氧条件下从干燥种子生长的能力为多细胞真核生物提供了一个难得的机会来研究由氧气供应触发的线粒体生物发生的各个阶段。研究了缺氧1 d后水稻线粒体的功能和蛋白质组学，并与有氧生长7 d的水稻幼苗进行了比较。与适应空气和空气生长的幼苗相比，在缺氧条件下生长的水稻胚囊和从胚囊中分离出来的线粒体呼吸非常缓慢。关键线粒体蛋白标记物的免疫检测、等电聚焦电泳（SDS-PAGE）绘制可溶性线粒体蛋白质组图，以及液相色谱-串联质谱法对线粒体蛋白进行鸟枪测序，均显示缺氧和空气适应样品中线粒体蛋白主要功能类别的模式相似。活性分析表明，幼苗在空气适应过程中，呼吸氧化酶活性显著升高。线粒体膜蛋白的蓝源电泳和SDS-PAGE显示，缺氧条件下线粒体膜中组装的b/c复合物和细胞色素c1氧化酶复合物丰度极低，而这些复合物的丰度在空气适应条件下急剧增加。总血红素含量、细胞色素吸收光谱和电子载体细胞色素c也显著增加。这些结果可能反映了在缺氧情况下细胞色素组装的血红素合成有限，并且在这种耐缺氧物种中代表了完全线粒体生物发生的离散和可逆阻塞。
The ability of rice seedlings to grow from dry seed under anoxia provides a rare opportunity in a multicellular eukaryote to study the stages of mitochondrial biogenesis triggered by oxygen availability. The function and proteome of rice mitochondria synthesized under 6 days of anoxia following 1 day of air adaptation have been compared with mitochondria isolated from 7-day aerobically grown rice seedlings. Rice coleoptiles grown under anoxia, and the mitochondria isolated from them respired very slowly compared with air-adapted and air-grown seedlings. Immunodetection of key mitochondrial protein markers, isoelectric focusing electrophoresis followed by SDS-PAGE to make soluble mitochondria proteome maps, and shotgun sequencing of mitochondrial proteins by liquid chromatography-tandem mass spectrometry all revealed similar patterns of the major function categories of mitochondrial proteins from both anoxic and air-adapted samples. Activity analysis showed respiratory oxidases markedly increased in activity during the air adaptation of seedlings. Blue-native electrophoresis followed by SDS-PAGE of mitochondrial membrane proteins clearly showed the very low abundance of assembled b/c complex and cytochrome c1 oxidase complex in the mitochondrial membrane in anoxic samples and the dramatic increase in the abundance of these complexes on air adaptation. Total heme content, cytochrome absorbance spectra, and the electron carrier, cytochrome c, also increased markedly on air adaptation. These results likely reflect limited heme synthesis for cytochrome assembly in the absence of oxygen and represent a discrete and reversible blockage of full mitochondrial biogenesis in this anoxia-tolerant species.