Imaging and Profiling of Proteins under Oxidative Conditions in Cells and Tissues by Hydrogen-Peroxide-Responsive Labeling
| Autor: | Yuki Nishikawa, Tomonori Tamura, Itaru Hamachi, Mikiko Takato, Hao Zhu, Rong Cheng, Alma Fujisawa |
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| Rok vydání: | 2020 |
| Předmět: |
Proteomics
Fluorescence-lifetime imaging microscopy Cell signaling Oxidative phosphorylation Mitochondrion 010402 general chemistry medicine.disease_cause 01 natural sciences Biochemistry Catalysis Mice Colloid and Surface Chemistry medicine Animals Fluorescent Dyes chemistry.chemical_classification Reactive oxygen species Dose-Response Relationship Drug Molecular Structure Proteomic Profiling Macrophages Optical Imaging General Chemistry Hydrogen Peroxide 0104 chemical sciences Cell biology Oxidative Stress RAW 264.7 Cells chemistry Oxidation-Reduction Oxidative stress |
| Zdroj: | Journal of the American Chemical Society. 142(37) |
| ISSN: | 1520-5126 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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