Sensitive detection of the redox state of copper proteins using fluorescence

The blue copper protein azurin from Pseudomonas aeruginosa has been covalently labelled with the fluorescing dye Cy5. The optical spectrum of the azurin changes markedly with its redox state. These changes are reflected in the fluorescence intensity of the dye through fluorescence resonance energy t...

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Bibliographic Details
Published in:Journal of biological inorganic chemistry 2005-10, Vol.10 (6), p.683-687
Main Authors: Schmauder, Ralf, Alagaratnam, Sharmini, Chan, Chris, Schmidt, Thomas, Canters, Gerard W, Aartsma, Thijs J
Format: Article
Language:English
Subjects:
Azurin - chemistry
Azurin - genetics
Carbocyanines - chemistry
Copper - chemistry
Fluorescence
Fluorescent Dyes - chemistry
Metalloproteins - chemistry
Mutation
Oxidation-Reduction
Pseudomonas aeruginosa - metabolism
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Summary:The blue copper protein azurin from Pseudomonas aeruginosa has been covalently labelled with the fluorescing dye Cy5. The optical spectrum of the azurin changes markedly with its redox state. These changes are reflected in the fluorescence intensity of the dye through fluorescence resonance energy transfer (FRET). This provides a sensitive way to monitor biological redox events. The method shown to work in the nanomolar range of protein concentrations, can be easily extended into the sub-nanomolar regime and holds promise for single-molecule detection.
ISSN:0949-8257
1432-1327
DOI:10.1007/s00775-005-0020-6