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Nat Commun. 2014 Oct 21;5:5222. doi: 10.1038/ncomms6222.

Red fluorescent genetically encoded indicator for intracellular hydrogen peroxide.

Author information

1
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia.
2
1] Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia [2] NBIC, Moscow Institute of Physics and Technology, 123182 Moscow, Russia.
3
NBIC, Moscow Institute of Physics and Technology, 123182 Moscow, Russia.
4
Department of Biophysics, School of Medicine, Saarland University, Homburg 66421, Germany.
5
1] NBIC, Moscow Institute of Physics and Technology, 123182 Moscow, Russia [2] Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA.

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 Ca(2+) uptake.

PMID:
25330925
PMCID:
PMC4553041
DOI:
10.1038/ncomms6222
[Indexed for MEDLINE]
Free PMC Article

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