Imaging Dynamic Redox Changes in Mammalian Cells with Green Fluorescent Protein Indicators*
By C. Dooley, Timothy M. Dore, George T. Hanson, W. C. Jackson, S. J. Remington, Roger Tsien. Journal of Biological Chemistry, 2004
733 citations recorded by the source corpus.
Changes in the redox equilibrium of cells influence a host of cell functions. Alterations in the redox equilibrium are precipitated by changing either the glutathione/glutathione-disulfide ratio (GSH/GSSG) and/or the reduced/oxidized thioredoxin ratio. Redox-sensitive green fluorescent proteins (GFP) allow real time visualization of the oxidation state of the indicator. Ratios of fluorescence from excitation at 400 and 490 nm indicate the extent of oxidation and thus the redox potential while canceling out the amount of indicator and the absolute optical sensitivity. Because the indicator is genetically encoded, it can be targeted to specific proteins or organelles of interest and expressed in a wide variety of cells and organisms. We evaluated roGFP1 (GFP with mutations C48S, S147C, and Q204C) and roGFP2 (the same plus S65T) with physiologically or toxicologically relevant oxidants both in vitro and in living mammalian cells. Furthermore, we investigated 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.
Subfields: Biophysics, Molecular Biology
View Paper: https://www.semanticscholar.org/paper/55255efd0012d5d3bca057107e3566a7934b4e5a
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Citations in the corpus
- Genetically encoded fluorescent indicator for intracellular hydrogen peroxide (2006, methodology): “Experiments that demonstrate temporal H 2 O 2 increase upon growth factor stimulation allowed comparison of HyPer with two other probes—H 2 DCFDA, the widely used dye for detection of intracellular ROS production, and roGFPs, recently developed redox-sensitive mutants of GF”
- Fluorescent proteins and their applications in imaging living cells and tissues. (2010, unknown): “value (147, 193, 216, 280, 286, 362), concentrations of chloride (190, 464) and metal ions (31, 246), and redox potential (94, 146).”
- Mitochondria-Ros Crosstalk in the Control of Cell Death and Aging (2011, unknown): “proteins (roGFPs) [103, 104].”