Hypocrates is a genetically encoded fluorescent biosensor for (pseudo)hypohalous acids and their derivatives

Alexander I. Kostyuk, Maria-Armineh Tossounian, Anastasiya S. Panova, Marion Thauvin, Roman I. Raevskii, Daria Ezeriņa, Khadija Wahni, Inge Molle, Anastasia D. Sergeeva, Didier Vertommen, Andrey Yu. Gorokhovatsky, Mikhail S. Baranov, Sophie Vriz, Joris Messens (), Dmitry S. Bilan () and Vsevolod V. Belousov ()
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Alexander I. Kostyuk: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Maria-Armineh Tossounian: VIB-VUB Center for Structural Biology, Vlaams Instituut voor Biotechnologie
Anastasiya S. Panova: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Marion Thauvin: INSERM, PSL Research University
Roman I. Raevskii: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Daria Ezeriņa: VIB-VUB Center for Structural Biology, Vlaams Instituut voor Biotechnologie
Khadija Wahni: VIB-VUB Center for Structural Biology, Vlaams Instituut voor Biotechnologie
Inge Molle: VIB-VUB Center for Structural Biology, Vlaams Instituut voor Biotechnologie
Anastasia D. Sergeeva: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Didier Vertommen: de Duve Institute, MASSPROT platform, UCLouvain
Andrey Yu. Gorokhovatsky: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Mikhail S. Baranov: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Sophie Vriz: INSERM, PSL Research University
Joris Messens: VIB-VUB Center for Structural Biology, Vlaams Instituut voor Biotechnologie
Dmitry S. Bilan: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Vsevolod V. Belousov: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry

Nature Communications, 2022, vol. 13, issue 1, 1-17

Abstract: Abstract The lack of tools to monitor the dynamics of (pseudo)hypohalous acids in live cells and tissues hinders a better understanding of inflammatory processes. Here we present a fluorescent genetically encoded biosensor, Hypocrates, for the visualization of (pseudo)hypohalous acids and their derivatives. Hypocrates consists of a circularly permuted yellow fluorescent protein integrated into the structure of the transcription repressor NemR from Escherichia coli. We show that Hypocrates is ratiometric, reversible, and responds to its analytes in the 106 M−1s−1 range. Solving the Hypocrates X-ray structure provided insights into its sensing mechanism, allowing determination of the spatial organization in this circularly permuted fluorescent protein-based redox probe. We exemplify its applicability by imaging hypohalous stress in bacteria phagocytosed by primary neutrophils. Finally, we demonstrate that Hypocrates can be utilized in combination with HyPerRed for the simultaneous visualization of (pseudo)hypohalous acids and hydrogen peroxide dynamics in a zebrafish tail fin injury model.

Date: 2022
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DOI: 10.1038/s41467-021-27796-2

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