Ultrasensitive Genetically Encoded Indicator for Hydrogen Peroxide Identifies Roles for the Oxidant in Cell Migration and Mitochondrial Function

We present HyPer7, the first bright, pH-stable, ultrafast, and ultrasensitive ratiometric H2O2 probe

Valeriy V. Pak; 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 V. Belousov

2020

Scholarcy highlights

  • Hydrogen peroxide is one of the key intermediates of aerobic metabolism
  • Addition of D-Ala induced a substantial increase of the HyPer7-intermembrane space signal, indicating that some part of the H2O2 produced in the matrix can leak to the IMS
  • Control experiments with H2O2-insensitive HyPer7-C121S demonstrated no change in the signal, confirming that the gradient observed with HyPer7 is due to the specific oxidation of the probe
  • We do not know the details of the redox environment of N. meningitidis, a cause of bacterial meningitis, one could suggest that it evolved the ultrasensitive OxyR to ensure a very early response to survive host immune cells
  • The explanations of the high reaction rate of N. meningitidis OxyR may lie in its active site pocket: while most OxyRs have a histidine residue before the peroxidatic cysteine, the OxyRs from the Neisseria group are the only ones with an asparagine residue at this position
  • V.V.P. and V.V.B. designed the research; B.P. performed OxyR multiple sequence analysis and phylogeny; V.V.P., D.E., D.Y., K.W., S.A.M.G., A.D.D., Y.D.M., D.S.B., and J.M. performed in vitro HyPer7 analysis; V.V.P., P.A.T.-K., Y.G.E., N.M.M., A.G.S., I.B., E.E., and T.M. performed live cell imaging and analysis; O.G.L. performed flow cytometry experiments and analysis; M.T., A.G.S., and S.V. performed experiments with zebrafish; V.V.B. wrote the manuscript with contributions from V.V.P., D.E., O.G.L., B.P., P.A.T.-K., T.M., S.V., and J.M.; V.V.B. supervised all aspects of the study; and all authors read the final version of the manuscript
  • HyPer7 is a probe of choice for realtime H2O2 imaging in various biological contexts

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