title: Development of redox-sensitive red fluorescent proteins for imaging redox dynamics in cellular compartments.
abstract: We recently reported a redox-sensitive red fluorescent protein, rxRFP1, which is one of the first genetically encoded red-fluorescent probes for general redox states in living cells. As individual cellular compartments have different basal redox potentials, we hereby describe a group of rxRFP1 mutants, showing different midpoint redox potentials for detection of redox dynamics in various subcellular domains, such as mitochondria, the cell nucleus, and endoplasmic reticulum (ER). When these redox probes were expressed and subcellularly localized in human embryonic kidney (HEK) 293 T cells, they responded to membrane-permeable oxidants and reductants. In addition, a mitochondrially localized rxRFP1 mutant, Mito-rxRFP1.1, was used to detect mitochondrial oxidative stress induced by doxorubicin-a widely used cancer chemotherapy drug. Our work has expanded the fluorescent protein toolkit with new research tools for studying compartmentalized redox dynamics and oxidative stress under various pathophysiological conditions.
authors.affiliation: Environmental Toxicology Graduate Program, University of California Riverside, 501 Big Springs Road, Riverside, CA, 92521, USA.
authors.forename: Yichong
authors.initials: Y
authors.lastname: Fan
authors.affiliation: Environmental Toxicology Graduate Program, University of California Riverside, 501 Big Springs Road, Riverside, CA, 92521, USA. huiwang.ai@ucr.edu.|Department of Chemistry, University of California Riverside, 501 Big Springs Road, Riverside, CA, 92521, USA. huiwang.ai@ucr.edu.
authors.forename: Hui-wang
authors.initials: HW
authors.lastname: Ai
mesh_terms: D000595:Amino Acid Sequence; D002451:Cell Compartmentation; D057809:HEK293 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry*; D008928:Mitochondria / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D017386:Sequence Homology, Amino Acid; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein
keywords: Compartmentalized redox dynamics; Fluorescence imaging; Fluorescent protein; Genetically encoded probe; Oxidative stress