Proteomic analysis of 4-hydroxynonenal (4-HNE) modified proteins in liver mitochondria from chronic ethanol-fed rats.
Proteomic analysis of 4-hydroxynonenal (4-HNE) modified proteins in liver mitochondria from chronic ethanol-fed rats.
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DOI：
10.1016/j.redox.2014.09.006
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发表时间：
2014
期刊：
影响因子：
11.4
通讯作者：
Bailey, Shannon M.
中科院分区：
文献类型：
--
作者：
Andringa, Kelly K.;Udoh, Uduak S.;Landar, Aimee;Bailey, Shannon M.
慢性乙醇介导的氧化应激和脂质过氧化增加了各种反应性脂质物质的水平，包括4-羟基壬烯醛（4-HNE），其随后可以修饰肝脏中的蛋白质。已经提出4-HNE修饰不利地影响线粒体蛋白的结构和/或功能，从而损害线粒体代谢。为了确定慢性乙醇消耗是否增加线粒体中4-HNE修饰蛋白的水平，雄性大鼠喂食对照和含乙醇饮食5周，并使用互补蛋白质组学方法分析线粒体样品。5条蛋白质条带（约35，45，50，70，和90 kDa）显示出强烈的免疫反应性4-HNE修饰的蛋白质在肝线粒体从对照组和乙醇喂养的大鼠时，蛋白质分离的标准一维SDS-PAGE。利用高分辨率蛋白质组学方法（2D IEF/SDS-PAGE和BN-PAGE），我们鉴定了几种与4-HNE免疫反应的线粒体蛋白，包括丝裂素、二甲基甘氨酸脱氢酶、胆碱脱氢酶、电子传递黄素蛋白α、细胞色素c1、烯酰CoA水合酶和细胞色素c。与对照组相比，电子转移黄素蛋白α在乙醇喂养大鼠的线粒体中始终显示4-HNE免疫反应性增加。增加的4-HNE反应性也检测到二甲基甘氨酸脱氢酶，烯酰CoA水合酶，和细胞色素c乙醇样品时，线粒体进行了BN-PAGE分析。总之，这项工作确定了线粒体中4-HNE修饰的新靶点，并提供了更好地理解慢性乙醇诱导的线粒体功能障碍和肝损伤的分子机制所需的有用信息。雄性大鼠喂食等热量Lieber-DeCarli对照和含乙醇饮食5周。分离线粒体并使用蛋白质组学方法检查4-HNE修饰。少数线粒体蛋白被发现是4-HNE的免疫反应。慢性乙醇消耗增加电子转移黄素蛋白-α中的4-HNE反应性。
Chronic ethanol-mediated oxidative stress and lipid peroxidation increases the levels of various reactive lipid species including 4-hydroxynonenal (4-HNE), which can subsequently modify proteins in the liver. It has been proposed that 4-HNE modification adversely affects the structure and/or function of mitochondrial proteins, thereby impairing mitochondrial metabolism. To determine whether chronic ethanol consumption increases levels of 4-HNE modified proteins in mitochondria, male rats were fed control and ethanol-containing diets for 5 weeks and mitochondrial samples were analyzed using complementary proteomic methods. Five protein bands (approx. 35, 45, 50, 70, and 90 kDa) showed strong immunoreactivity for 4-HNE modified proteins in liver mitochondria from control and ethanol-fed rats when proteins were separated by standard 1D SDS-PAGE. Using high-resolution proteomic methods (2D IEF/SDS-PAGE and BN-PAGE) we identified several mitochondrial proteins immunoreactive for 4-HNE, which included mitofilin, dimethylglycine dehydrogenase, choline dehydrogenase, electron transfer flavoprotein α, cytochrome c1, enoyl CoA hydratase, and cytochrome c. The electron transfer flavoprotein α consistently showed increased 4-HNE immunoreactivity in mitochondria from ethanol-fed rats as compared to mitochondria from the control group. Increased 4-HNE reactivity was also detected for dimethylglycine dehydrogenase, enoyl CoA hydratase, and cytochrome c in ethanol samples when mitochondria were analyzed by BN-PAGE. In summary, this work identifies new targets of 4-HNE modification in mitochondria and provides useful information needed to better understand the molecular mechanisms underpinning chronic ethanol-induced mitochondrial dysfunction and liver injury. Male rats were fed isocaloric Lieber–DeCarli control and ethanol-containing diets for 5 weeks. Mitochondria were isolated and examined for 4-HNE modifications using proteomic methods. A small number of mitochondrial proteins were found to be immunoreactive for 4-HNE. Chronic ethanol consumption increased 4-HNE reactivity in the electron transfer flavoprotein-α.
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影响因子：
5.1
作者：
Smathers, Rebecca L.;Galligan, James J.;Stewart, Benjamin J.;Petersen, Dennis R.
通讯作者：
Petersen, Dennis R.
DOI：
10.1152/ajpgi.00044.2006
发表时间：
2006-11-01
影响因子：
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作者：
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10.1124/jpet.105.088088
发表时间：
2005-10-01
影响因子：
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影响因子：
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影响因子：
13.5
作者：
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