PUBLICATION
Hypocrates is a genetically encoded fluorescent biosensor for (pseudo)hypohalous acids and their derivatives
- Authors
- Kostyuk, A.I., Tossounian, M.A., Panova, A.S., Thauvin, M., Raevskii, R.I., Ezeriņa, D., Wahni, K., Van Molle, I., Sergeeva, A.D., Vertommen, D., Gorokhovatsky, A.Y., Baranov, M.S., Vriz, S., Messens, J., Bilan, D.S., Belousov, V.V.
- ID
- ZDB-PUB-220112-8
- Date
- 2022
- Source
- Nature communications 13: 171 (Journal)
- Registered Authors
- Vriz, Sophie
- Keywords
- none
- MeSH Terms
-
- Transcription Factors/genetics
- Transcription Factors/metabolism
- Fluorescent Dyes/chemistry*
- Animals
- Escherichia coli/genetics
- Escherichia coli/metabolism
- Genes, Reporter
- Zebrafish
- Biosensing Techniques/instrumentation
- Biosensing Techniques/methods*
- Animal Fins/diagnostic imaging*
- Animal Fins/injuries
- Animal Fins/metabolism
- Crystallography, X-Ray
- Hypochlorous Acid/analysis*
- Hypochlorous Acid/chemical synthesis
- Hypochlorous Acid/metabolism
- Escherichia coli Proteins/genetics
- Escherichia coli Proteins/metabolism
- Neutrophils/cytology
- Neutrophils/immunology
- Luminescent Proteins/genetics*
- Luminescent Proteins/metabolism
- Hydrogen Peroxide/chemistry
- Oxidation-Reduction
- Phagocytosis
- Bacterial Proteins/genetics*
- Bacterial Proteins/metabolism
- PubMed
- 35013284 Full text @ Nat. Commun.
Citation
Kostyuk, A.I., Tossounian, M.A., Panova, A.S., Thauvin, M., Raevskii, R.I., Ezeriņa, D., Wahni, K., Van Molle, I., Sergeeva, A.D., Vertommen, D., Gorokhovatsky, A.Y., Baranov, M.S., Vriz, S., Messens, J., Bilan, D.S., Belousov, V.V. (2022) Hypocrates is a genetically encoded fluorescent biosensor for (pseudo)hypohalous acids and their derivatives. Nature communications. 13:171.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping