Ultrasensitive Genetically Encoded Indicator for Hydrogen Peroxide Identifies Roles for the Oxidant in Cell Migration and Mitochondrial Function.
Citation | Pak, Valeriy , V, et al. “Ultrasensitive Genetically Encoded Indicator for Hydrogen Peroxide Identifies Roles for the Oxidant in Cell Migration and Mitochondrial Function”. 2020. Cell Metabolism, vol. 31, no. 3, 2020, pp. 642–653.e6. |
Center | Boston Area |
Author | Valeriy Pak V, Daria Ezeriņa, Olga G Lyublinskaya, Brandán Pedre, Pyotr A Tyurin-Kuzmin, Natalie M Mishina, Marion Thauvin, David Young, Khadija Wahni, Santiago Agustín Martínez Gache, Alexandra D Demidovich, Yulia G Ermakova, Yulia D Maslova, Arina G Shokhina, Emrah Eroglu, Dmitry S Bilan, Ivan Bogeski, Thomas Michel, Sophie Vriz, Joris Messens, Vsevolod Belousov V |
Keywords | D-amino acid oxidase, H(2)O(2), H(2)O(2) gradients, HyPer7, cell migration, chemogenetics, genetically encoded probes, Hydrogen peroxide, mitochondria, Redox signaling |
Abstract |
Hydrogen peroxide (HO) is a key redox intermediate generated within cells. Existing probes for HO have not solved the problem of detection of the ultra-low concentrations of the oxidant: these reporters are not sensitive enough, or pH-dependent, or insufficiently bright, or not functional in mammalian cells, or have poor dynamic range. Here we present HyPer7, the first bright, pH-stable, ultrafast, and ultrasensitive ratiometric HO probe. HyPer7 is fully functional in mammalian cells and in other higher eukaryotes. The probe consists of a circularly permuted GFP integrated into the ultrasensitive OxyR domain from Neisseria meningitidis. Using HyPer7, we were able to uncover the details of HO diffusion from the mitochondrial matrix, to find a functional output of HO gradients in polarized cells, and to prove the existence of HO gradients in wounded tissue in vivo. Overall, HyPer7 is a probe of choice for real-time HO imaging in various biological contexts. |
Year of Publication |
2020
|
Journal |
Cell metabolism
|
Volume |
31
|
Issue |
3
|
Number of Pages |
642-653.e6
|
Date Published |
12/2020
|
ISSN Number |
1932-7420
|
DOI |
10.1016/j.cmet.2020.02.003
|
Alternate Journal |
Cell Metab
|
PMID |
32130885
|
PMCID |
PMC7088435
|
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