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
  • Animal Fins/diagnostic imaging*
  • Animal Fins/injuries
  • Animal Fins/metabolism
  • Animals
  • Bacterial Proteins/genetics*
  • Bacterial Proteins/metabolism
  • Biosensing Techniques/instrumentation
  • Biosensing Techniques/methods*
  • Crystallography, X-Ray
  • Escherichia coli/genetics
  • Escherichia coli/metabolism
  • Escherichia coli Proteins/genetics
  • Escherichia coli Proteins/metabolism
  • Fluorescent Dyes/chemistry*
  • Genes, Reporter
  • Hydrogen Peroxide/chemistry
  • Hypochlorous Acid/analysis*
  • Hypochlorous Acid/chemical synthesis
  • Hypochlorous Acid/metabolism
  • Luminescent Proteins/genetics*
  • Luminescent Proteins/metabolism
  • Neutrophils/cytology
  • Neutrophils/immunology
  • Oxidation-Reduction
  • Phagocytosis
  • Transcription Factors/genetics
  • Transcription Factors/metabolism
  • Zebrafish
PubMed
35013284 Full text @ Nat. Commun.
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
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping