Intracellular distribution of oxidized proteins and proteasome in HT22 cells during oxidative stress
Intracellular distribution of oxidized proteins and proteasome in HT22 cells during oxidative stress
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DOI：
10.1016/j.freeradbiomed.2005.11.023
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发表时间：
2006-04-15
影响因子：
7.4
通讯作者：
Grune, T
中科院分区：
文献类型：
--
作者：
Jung, T;Engels, M;Grune, T
自由基的产生和由此导致的细胞结构的氧化损伤总是与氧化蛋白质的形成有关。20S蛋白酶体负责识别和降解氧化损伤的蛋白质。到目前为止，还没有关于氧化应激过程中氧化蛋白在细胞内的分布以及蛋白酶体的分布的详细研究。因此，我们使用免疫化学方法来测量蛋白质羰基，一种蛋白质氧化产物的形式，以及蛋白酶体在细胞内的分布。这两种免疫细胞化学方法都是半定量的，在探索了各种氧化应激后，氧化蛋白的负荷增加了，其中以细胞核周区域的增加最高。蛋白酶体的分布和总蛋白含量显示两者在细胞核中的浓度最高。氧化应激过程中不会发生蛋白酶体的重新分布。蛋白质羰基与蛋白质含量的归一化比值形成，表明氧化蛋白质在靠近细胞膜的胞液区域浓度最高。通过形成蛋白质氧化与蛋白酶体的比率，可以得出结论，氧化蛋白进入蛋白酶体的最大负荷发生在胞浆中，与所研究的氧化剂无关。(C)2005 Elsevier Inc.保留所有权利。
The production of free radicals and the resulting oxidative damage of cellular structures are always connected with the formation of oxidized proteins. The 20S proteasome is responsible for recognition and degradation of oxidatively damaged proteins. No detailed studies on the intracellular distribution of oxidized proteins during oxidative stress and on the distribution of the proteasome have been performed until now. Therefore, we used immunochemical methods to measure protein carbonyls, a form of protein oxidation products, and proteasome distribution within cells. Both immunocytochemical methods of measurement are semiquantitative and the load of oxidized proteins is increased after various oxidative stresses explored, with the highest increase in the perinuclear region of the cell. Distribution of the proteasome and the total protein content revealed the highest concentration of both in the nucleus. No redistribution of the proteasome during oxidative stress Occurs. The normalized ratio of protein carbonyls to protein content was formed, indicating the highest concentration of oxidized proteins in the cytosolic region near the cell membrane. By forming the protein oxidation-to-proteasome ratio it was concluded that the highest load of oxidized proteins to the proteasome takes place in the cytosol, independent of the oxidant explored. (c) 2005 Elsevier Inc. All rights reserved.