Red fluorescent genetically encoded indicator for intracellular hydrogen peroxide

• Published in: Nature communications Vol. 5; no. 1; p. 5222
• Main Authors: Ermakova, Yulia G., Bilan, Dmitry S., Matlashov, Mikhail E., Mishina, Natalia M., Markvicheva, Ksenia N., Subach, Oksana M., Subach, Fedor V., Bogeski, Ivan, Hoth, Markus, Enikolopov, Grigori, Belousov, Vsevolod V.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 21-10-2014; Nature Publishing Group
• Subjects: 14; 14/10; 14/35; 14/63; 631/1647/1888; 631/45; Calcium - chemistry; Cytoplasm - chemistry; Electron Transport; Fluorescent Dyes - chemistry; HEK293 Cells; HeLa Cells; Humanities and Social Sciences; Humans; Hydrogen Peroxide - chemistry; Kinetics; Luminescent Proteins - chemistry; Microscopy, Fluorescence; Mitochondria - metabolism; multidisciplinary; Oxidation-Reduction; Reactive Oxygen Species - chemistry; Recombinant Proteins - chemistry; Red Fluorescent Protein; Science; Science (multidisciplinary); Signal Transduction; Spectrophotometry; Time Factors
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms6222

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 H
2
O
2
produced in the cytoplasm of cultured cells upon growth factor stimulation. Moreover, using HyPerRed we detect local and transient H
2
O
2
production in the mitochondrial matrix upon inhibition of the endoplasmic reticulum Ca
2+
uptake.
Current genetically encoded sensors for hydrogen peroxide, such as HyPer, emit in the green region giving potential for overlap with other probes. Here, the authors report HyPerRed, a red fluorescent redox probe capable of monitoring intracellular hydrogen peroxide.