Imaging and profiling of proteins under oxidative conditions in cells and tissues by hydrogen-peroxide-responsive labeling,Journal of the American Chemical Society

当前位置： X-MOL 学术 › J. Am. Chem. Soc. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Imaging and profiling of proteins under oxidative conditions in cells and tissues by hydrogen-peroxide-responsive labeling
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-08-21 , DOI: 10.1021/jacs.0c02547
Hao Zhu ₁ , Tomonori Tamura _{1,

2} , Alma Fujisawa ₁ , Yuki Nishikawa ₁ , Rong Cheng ₁ , Mikiko Takato ₁ , Itaru Hamachi _{1,

2}

Affiliation

Reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) can inflict damage to biomolecules under oxidative stress and also act as signaling molecules at physiological levels. Here we developed a unique chemical tool to elucidate the biological roles of ROS using both fluorescence imaging and conditional proteomics. H2O2-responsive protein labeling reagents (Hyp-L) were designed to selectively tag proteins under the oxidative conditions in living cells and tissues. The Hyp-L signal remained even after sample fixation, which was compatible with conventional immunostaining. Moreover, Hyp-L allowed proteomic profiling of the labeled proteins using a conditional proteomics workflow. The integrative analysis enabled the identification of ROS generation and/or accumulation sites with a subcellular resolution. For the first time, we characterized that autophagosomes were enriched with H2O2 in activated macrophages. Hyp-L was further applied to mouse brain tissues and clearly revealed oxidative stress within mitochondria by the conditional proteomics.

中文翻译：

通过过氧化氢反应标记对细胞和组织中氧化条件下的蛋白质进行成像和分析

过氧化氢 (H2O2) 等活性氧 (ROS) 可在氧化应激下对生物分子造成损害，并在生理水平上充当信号分子。在这里，我们开发了一种独特的化学工具，使用荧光成像和条件蛋白质组学来阐明 ROS 的生物学作用。H2O2 反应性蛋白质标记试剂 (Hyp-L) 旨在在活细胞和组织中的氧化条件下选择性标记蛋白质。即使在样品固定后，Hyp-L 信号仍然存在，这与常规免疫染色兼容。此外，Hyp-L 允许使用条件蛋白质组学工作流程对标记蛋白质进行蛋白质组学分析。综合分析能够以亚细胞分辨率识别 ROS 生成和/或积累位点。首次，我们的特征是自噬体在活化的巨噬细胞中富含 H2O2。Hyp-L 进一步应用于小鼠脑组织，并通过条件蛋白质组学清楚地揭示了线粒体内的氧化应激。

更新日期：2020-08-21

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11