Using the heme peroxidase APEX2 to probe intracellular H2O2 flux and diffusion
Abstract Currently available genetically encoded H2O2 probes report on the thiol redox state of the probe, which means that they reflect the balance between probe thiol oxidation and reduction. Here we introduce the use of the engineered heme peroxidase APEX2 as a thiol-independent chemogenetic H2O2...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-02-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-45511-9 |
| _version_ | 1827326931805143040 |
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| author | Mohammad Eid Uladzimir Barayeu Kateřina Sulková Carla Aranda-Vallejo Tobias P. Dick |
| author_facet | Mohammad Eid Uladzimir Barayeu Kateřina Sulková Carla Aranda-Vallejo Tobias P. Dick |
| author_sort | Mohammad Eid |
| collection | DOAJ |
| description | Abstract Currently available genetically encoded H2O2 probes report on the thiol redox state of the probe, which means that they reflect the balance between probe thiol oxidation and reduction. Here we introduce the use of the engineered heme peroxidase APEX2 as a thiol-independent chemogenetic H2O2 probe that directly and irreversibly converts H2O2 molecules into either fluorescent or luminescent signals. We demonstrate sensitivity, specificity, and the ability to quantitate endogenous H2O2 turnover. We show how the probe can be used to detect changes in endogenous H2O2 generation and to assess the roles and relative contributions of endogenous H2O2 scavengers. Furthermore, APEX2 can be used to study H2O2 diffusion inside the cytosol. Finally, APEX2 reveals the impact of commonly used alkylating agents and cell lysis protocols on cellular H2O2 generation. |
| first_indexed | 2024-03-07T14:52:39Z |
| format | Article |
| id | doaj.art-8293dadb5d7a4ec5b531d595301109e2 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-07T14:52:39Z |
| publishDate | 2024-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-8293dadb5d7a4ec5b531d595301109e22024-03-05T19:35:39ZengNature PortfolioNature Communications2041-17232024-02-0115111510.1038/s41467-024-45511-9Using the heme peroxidase APEX2 to probe intracellular H2O2 flux and diffusionMohammad Eid0Uladzimir Barayeu1Kateřina Sulková2Carla Aranda-Vallejo3Tobias P. Dick4Division of Redox Regulation, DKFZ-ZMBH Alliance, German Cancer Research Center (DKFZ)Division of Redox Regulation, DKFZ-ZMBH Alliance, German Cancer Research Center (DKFZ)Division of Redox Regulation, DKFZ-ZMBH Alliance, German Cancer Research Center (DKFZ)Division of Redox Regulation, DKFZ-ZMBH Alliance, German Cancer Research Center (DKFZ)Division of Redox Regulation, DKFZ-ZMBH Alliance, German Cancer Research Center (DKFZ)Abstract Currently available genetically encoded H2O2 probes report on the thiol redox state of the probe, which means that they reflect the balance between probe thiol oxidation and reduction. Here we introduce the use of the engineered heme peroxidase APEX2 as a thiol-independent chemogenetic H2O2 probe that directly and irreversibly converts H2O2 molecules into either fluorescent or luminescent signals. We demonstrate sensitivity, specificity, and the ability to quantitate endogenous H2O2 turnover. We show how the probe can be used to detect changes in endogenous H2O2 generation and to assess the roles and relative contributions of endogenous H2O2 scavengers. Furthermore, APEX2 can be used to study H2O2 diffusion inside the cytosol. Finally, APEX2 reveals the impact of commonly used alkylating agents and cell lysis protocols on cellular H2O2 generation.https://doi.org/10.1038/s41467-024-45511-9 |
| spellingShingle | Mohammad Eid Uladzimir Barayeu Kateřina Sulková Carla Aranda-Vallejo Tobias P. Dick Using the heme peroxidase APEX2 to probe intracellular H2O2 flux and diffusion Nature Communications |
| title | Using the heme peroxidase APEX2 to probe intracellular H2O2 flux and diffusion |
| title_full | Using the heme peroxidase APEX2 to probe intracellular H2O2 flux and diffusion |
| title_fullStr | Using the heme peroxidase APEX2 to probe intracellular H2O2 flux and diffusion |
| title_full_unstemmed | Using the heme peroxidase APEX2 to probe intracellular H2O2 flux and diffusion |
| title_short | Using the heme peroxidase APEX2 to probe intracellular H2O2 flux and diffusion |
| title_sort | using the heme peroxidase apex2 to probe intracellular h2o2 flux and diffusion |
| url | https://doi.org/10.1038/s41467-024-45511-9 |
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