High-Resolution Imaging of Redox Signaling in Live Cells Through an Oxidation-Sensitive Yellow Fluorescent Protein

  title={High-Resolution Imaging of Redox Signaling in Live Cells Through an Oxidation-Sensitive Yellow Fluorescent Protein},
  author={Giuseppe Maulucci and Valentina Labate and Marina Mele and Emiliano Panieri and Giuseppe Arc{\`o}vito and Tommaso Galeotti and Henrik {\O}stergaard and Jakob Rahr Winther and Marco De Spirito and Giovambattista Pani},
  journal={Science Signaling},
  pages={pl3 - pl3}
Measure and visualize intracellular changes in redox state. We present the application of a redox-sensitive mutant of the yellow fluorescent protein (rxYFP) to image, with elevated sensitivity and high temporal and spatial resolution, oxidative responses of eukaryotic cells to pathophysiological stimuli. The method presented, based on the ratiometric quantitation of the distribution of fluorescence by confocal microscopy, allows us to draw real-time “redox maps” of adherent cells and to score… 

Fluorescent protein-based redox probes.

Genetically encoded redox probes enable the functional analysis of individual proteins in cellular redox homeostasis, and redox biosensor transgenic model organisms offer extended opportunities for dynamic in vivo imaging of redox processes.

Dissecting Redox Biology Using Fluorescent Protein Sensors.

The growing range of thiol redox sensor variants and their application in different cells, tissues, and organisms is reviewed and how novel sensor variants may further add to the current momentum toward a novel mechanistic and integrated understanding of redox biology in vivo is explored.

Genetically encoded fluorescent redox sensors.

Genetically encoded reactive oxygen species (ROS) and redox indicators

  • S. Pouvreau
  • Environmental Science, Biology
    Biotechnology journal
  • 2014
This review is intended to be a tour guide of the main types of GERRIs, their origins, properties, advantages and pitfalls.



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It is demonstrated that the fusion of human glutaredoxin-1 to roGFP2 facilitates specific real-time equilibration between the sensor protein and the glutathione redox couple, which facilitated the observation of redox changes associated with growth factor availability, cell density, mitochondrial depolarization, respiratory burst activity and immune receptor stimulation.

Imaging Dynamic Redox Changes in Mammalian Cells with Green Fluorescent Protein Indicators*

The response of the redox probes under physiological redox changes during superoxide bursts in macrophage cells, hyperoxic and hypoxic conditions, and in responses to H2O2-stimulating agents, e.g. epidermal growth factor and lysophosphatidic acid is investigated.

Investigating Mitochondrial Redox Potential with Redox-sensitive Green Fluorescent Protein Indicators*

By substitution of surface-exposed residues on the Aequorea victoria green fluorescent protein with cysteines in appropriate positions to form disulfide bonds, reduction-oxidation-sensitive GFPs (roGFPs) have been created and targeted to the mitochondria in HeLa cells.

Measuring intracellular redox conditions using GFP-based sensors.

The redox sensors have been characterized biochemically and found to behave differently, both spectroscopically and in terms of redox properties; however, which parameters are measured depends on how the sensors interact with various cellular redox components.

Creating new fluorescent probes for cell biology

Advances include the continued development of 'passive' markers for the measurement of biomolecule expression and localization in live cells, and 'active' indicators for monitoring more complex cellular processes such as small-molecule-messenger dynamics, enzyme activation and protein–protein interactions.

Shedding light on disulfide bond formation: engineering a redox switch in green fluorescent protein

In the cytoplasm of Escherichia coli, the protein was a sensitive probe for the redox changes that occur upon disruption of the thioredoxin reductive pathway and a detailed mechanism accounting for the observed redox state‐dependent fluorescence is proposed.

Cell Compartmentalization in Redox Signaling

Experimental data supporting the concept that cell responses to redox changes are function of the intracellular site where oxidants are produced and/or meet their molecular targets are reviewed.

Redox-dependent transcriptional regulation.

This review summarizes the recent progress on how cellular redox status can regulate transcription-factor activity and the implications of this regulation for cardiovascular disease.