Estradiol In Vivo regulation of brain mitochondrial proteome
Estradiol In Vivo regulation of brain mitochondrial proteome
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DOI：
10.1523/jneurosci.4391-07.2007
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发表时间：
2007-12-19
影响因子：
5.3
通讯作者：
Brinton, Roberta Diaz
中科院分区：
文献类型：
--
作者：
Nilsen, Jon;Irwin, Ronald W.;Brinton, Roberta Diaz
我们使用蛋白质组学和功能生化相结合的方法来确定17 β-雌二醇（E-2）对线粒体蛋白表达和功能的总体影响。为了阐明E-2激活的脑线粒体通路，采用二维凝胶电泳技术筛选线粒体蛋白质组。切除卵巢的成年雌性大鼠单次注射E-2。在E-2暴露24 h后，从脑中纯化线粒体，并进行二维分析和液相色谱-串联质谱蛋白鉴定。蛋白质组学分析的结果表明，在通过图像分析检测到的499个蛋白质点中，共有66个蛋白质点的表达变化为2倍或更大。其中，28种蛋白质在E-2处理后表达增加，而38种蛋白质相对于对照表达减少。E-2调节关键代谢酶，包括丙酮酸脱氢酶、乌头酸酶和ATP合酶。为了证实E-2诱导的蛋白质表达变化转化为功能后果，我们确定了E-2对线粒体电子传递链酶活性的影响。在体内，E-2处理增强脑线粒体效率，如通过增加的呼吸控制比、升高的细胞色素-c氧化酶活性和表达所证明的，同时减少脑中自由基的产生。这些分析的结果提供了对调节脑线粒体的E-2机制的见解，其具有维持神经系统健康和预防与线粒体功能障碍相关的神经退行性疾病（如阿尔茨海默病）的潜力。
We used a combined proteomic and functional biochemical approach to determine the overall impact of 17 beta-estradiol (E-2) on mitochondrial protein expression and function. To elucidate mitochondrial pathways activated by E-2 in brain, two-dimensional (2D) gel electrophoresis was conducted to screen the mitoproteome. Ovariectomized adult female rats were treated with a single injection of E-2. After 24 h of E-2 exposure, mitochondria were purified from brain and 2D analysis and liquid chromatography-tandem mass spectrometry protein identification were conducted. Results of proteomic analyses indicated that of the 499 protein spots detected by image analysis, a total of 66 protein spots had a twofold or greater change in expression. Of these, 28 proteins were increased in expression after E-2 treatment whereas 38 proteins were decreased in expression relative to control. E-2 regulated key metabolic enzymes including pyruvate dehydrogenase, aconitase, and ATP-synthase. To confirm that E-2-inducible changes in protein expression translated into functional consequences, we determined the impact of E-2 on the enzymatic activity of the mitochondrial electron transport chain. In vivo, E-2 treatment enhanced brain mitochondrial efficiency as evidenced by increased respiratory control ratio, elevated cytochrome-c oxidase activity and expression while simultaneously reducing free radical generation in brain. Results of these analyses provide insights into E-2 mechanisms of regulating brain mitochondria, which have the potential for sustaining neurological health and prevention of neurodegenerative diseases associated with mitochondrial dysfunction such as Alzheimer's disease.