title: Expression and characterization of a redox-sensing green fluorescent protein (reduction-oxidation-sensitive green fluorescent protein) in Arabidopsis.
abstract: Arabidopsis (Arabidopsis thaliana) was transformed with a redox-sensing green fluorescent protein (reduction-oxidation-sensitive green fluorescent protein [roGFP]), with expression targeted to either the cytoplasm or to the mitochondria. Both the mitochondrial and cytosolic forms are oxidation-reduction sensitive, as indicated by a change in the ratio of 510 nm light (green light) emitted following alternating illumination with 410 and 474 nm light. The 410/474 fluorescence ratio is related to the redox potential (in millivolts) of the organelle, cell, or tissue. Both forms of roGFP can be reduced with dithiothreitol and oxidized with hydrogen peroxide. The average resting redox potentials for roots are -318 mV for the cytoplasm and -362 mV for the mitochondria. The elongation zone of the Arabidopsis root has a more oxidized redox status than either the root cap or meristem. Mitochondria are much better than the cytoplasm, as a whole, at buffering changes in redox. The data show that roGFP is redox sensitive in plant cells and that this sensor makes it possible to monitor, in real time, dynamic changes in redox in vivo.
authors.affiliation: Department of Plant and Microbial Biology , University of California, Berkeley, 94720, USA.
authors.forename: Keni
authors.initials: K
authors.lastname: Jiang
authors.forename: Christian
authors.initials: C
authors.lastname: Schwarzer
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Lally
authors.forename: Shibo
authors.initials: S
authors.lastname: Zhang
authors.forename: Steven
authors.initials: S
authors.lastname: Ruzin
authors.forename: Terry
authors.initials: T
authors.lastname: Machen
authors.forename: S James
authors.initials: SJ
authors.lastname: Remington
authors.forename: Lewis
authors.initials: L
authors.lastname: Feldman
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D001483:Base Sequence; D003593:Cytoplasm / Q000378:metabolism; D017931:DNA Primers; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D010084:Oxidation-Reduction
chemical_list: D017931:DNA Primers; D049452:Green Fluorescent Proteins