Red fluorescent genetically encoded indicator for intracellular hydrogen peroxide

Yulia G. Ermakova, Dmitry S. Bilan, Mikhail E. Matlashov, Natalia M. Mishina, Ksenia N. Markvicheva, Oksana M. Subach, Fedor V. Subach, Ivan Bogeski, Markus Hoth, Grigori Enikolopov () and Vsevolod V. Belousov ()
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Yulia G. Ermakova: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Dmitry S. Bilan: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Mikhail E. Matlashov: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Natalia M. Mishina: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Ksenia N. Markvicheva: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
Oksana M. Subach: NBIC, Moscow Institute of Physics and Technology
Fedor V. Subach: NBIC, Moscow Institute of Physics and Technology
Ivan Bogeski: School of Medicine, Saarland University
Markus Hoth: School of Medicine, Saarland University
Grigori Enikolopov: NBIC, Moscow Institute of Physics and Technology
Vsevolod V. Belousov: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry

Nature Communications, 2014, vol. 5, issue 1, 1-9

Abstract: Abstract Reactive oxygen species (ROS) are conserved regulators of numerous cellular functions, and overproduction of ROS is a hallmark of various pathological processes. Genetically encoded fluorescent probes are unique tools to study ROS production in living systems of different scale and complexity. However, the currently available recombinant redox sensors have green emission, which overlaps with the spectra of many other probes. Expanding the spectral range of recombinant in vivo ROS probes would enable multiparametric in vivo ROS detection. Here we present the first genetically encoded red fluorescent sensor for hydrogen peroxide detection, HyPerRed. The performance of this sensor is similar to its green analogues. We demonstrate the utility of the sensor by tracing low concentrations of H2O2 produced in the cytoplasm of cultured cells upon growth factor stimulation. Moreover, using HyPerRed we detect local and transient H2O2 production in the mitochondrial matrix upon inhibition of the endoplasmic reticulum Ca2+ uptake.

Date: 2014
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DOI: 10.1038/ncomms6222

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