title: A Förster-resonance-energy transfer-based method for fluorescence detection of the protein redox state.
abstract: A method for fluorescence detection of a protein's redox state based on resonance energy transfer from an attached fluorescence label to the prosthetic group of the redox protein is described and tested for proteins containing three types of prosthetic groups: a type-1 copper site (azurin, amicyanin, plastocyanin, and pseudoazurin), a heme group (cytochrome c550), and a flavin mononucleotide (flavodoxin). This method permits one to reliably distinguish between reduced and oxidized proteins and to perform potentiometric titrations at submicromolar concentrations.
authors.affiliation: Leiden Institute of Chemistry-Gorlaeus Laboratories, Leiden University, The Netherlands.
authors.forename: Sofya
authors.initials: S
authors.lastname: Kuznetsova
authors.forename: Gerhild
authors.initials: G
authors.lastname: Zauner
authors.forename: Ralf
authors.initials: R
authors.lastname: Schmauder
authors.forename: Oleg A
authors.initials: OA
authors.lastname: Mayboroda
authors.forename: André M
authors.initials: AM
authors.lastname: Deelder
authors.forename: Thijs J
authors.initials: TJ
authors.lastname: Aartsma
authors.forename: Gerard W
authors.initials: GW
authors.lastname: Canters
mesh_terms: D001400:Azurin / Q000737:chemistry; D002232:Carbocyanines / Q000737:chemistry; D003300:Copper / Q000737:chemistry; D003574:Cytochrome c Group / Q000737:chemistry; D005418:Flavodoxin / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D010084:Oxidation-Reduction; D011199:Potentiometry; D011506:Proteins / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D002232:Carbocyanines; D003574:Cytochrome c Group; D005418:Flavodoxin; D005456:Fluorescent Dyes; D011506:Proteins; C085321:cyanine dye 5; D001400:Azurin; D003300:Copper; C031732:cytochrome C-550