A DNA-based fluorescent reporter maps HOCl production in the maturing phagosome
| Autor: | Yamuna Krishnan, Guolin Zhou, Maulik S Jani, Shareefa Thekkan, Lev Becker, Chang Cui, Krishna Dan |
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| Rok vydání: | 2018 |
| Předmět: |
chemistry.chemical_classification
0303 health sciences Reactive oxygen species Hypochlorous acid Phagosome acidification 030302 biochemistry & molecular biology DNA Cell Biology Hydrogen-Ion Concentration Hypochlorous Acid Cell biology 03 medical and health sciences chemistry.chemical_compound chemistry Phagosomes Organelle Phagosome maturation Oxidation-Reduction Molecular Biology Reactive nitrogen species Fluorescent Dyes 030304 developmental biology Phagosome |
| Zdroj: | Nature Chemical Biology. 15:1165-1172 |
| ISSN: | 1552-4469 1552-4450 |
| DOI: | 10.1038/s41589-018-0176-3 |
| Popis: | Phagocytes destroy pathogens by trapping them in a transient organelle called the phagosome, where they are bombarded with reactive oxygen species (ROS) and reactive nitrogen species (RNS). Imaging reactive species within the phagosome would directly reveal the chemical dynamics underlying pathogen destruction. Here we introduce a fluorescent, DNA-based combination reporter, cHOClate, which simultaneously images hypochlorous acid (HOCl) and pH quantitatively. Using cHOClate targeted to phagosomes in live cells, we successfully map phagosomal production of a specific ROS, HOCl, as a function of phagosome maturation. We found that phagosomal acidification was gradual in macrophages and upon completion, HOCl was released in a burst. This revealed that phagosome-lysosome fusion was essential not only for phagosome acidification, but also for providing the chloride necessary for myeloperoxidase activity. This method can be expanded to image several kinds of ROS and RNS and be readily applied to identify how resistant pathogens evade phagosomal killing. |
| Databáze: | OpenAIRE |
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