An insight into pH-induced changes in FAD conformational structure by means of time-resolved fluorescence and circular dichroism

Optical properties of flavin adenine dinucleotide (FAD) moiety are widely used nowadays for biotechnological applications. Given the fundamental role played by FAD, additional structural information about this enzymatic cofactor can be extremely useful in order to obtain a greater insight into its f...

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Bibliographic Details
Published in:European biophysics journal 2019-05, Vol.48 (4), p.395-403
Main Authors: Esposito, Rosario, Delfino, Ines, Portaccio, Marianna, Iannuzzi, Clara, Lepore, Maria
Format: Article
Language:English
Subjects:
Aqueous solutions
Biochemistry
Biological and Medical Physics
Biomedical and Life Sciences
Biophysics
Cell Biology
Circular Dichroism
Dichroism
Emission analysis
Flavin-adenine dinucleotide
Flavin-Adenine Dinucleotide - chemistry
Fluorescence
Hydrogen-Ion Concentration
Life Sciences
Membrane Biology
Molecular Conformation
Nanotechnology
Neurobiology
Optical properties
Optics
Original Article
pH effects
Spectrometry, Fluorescence
Time Factors
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Summary:Optical properties of flavin adenine dinucleotide (FAD) moiety are widely used nowadays for biotechnological applications. Given the fundamental role played by FAD, additional structural information about this enzymatic cofactor can be extremely useful in order to obtain a greater insight into its functional role in proteins. For this purpose, we have investigated FAD behaviour in aqueous solutions at different pH values by a novel approach based on the combined use of time-resolved fluorescence and circular dichroism spectroscopies. The results showed that pH strongly affects time-resolved fluorescence emission and the analysis allowed us to detect a three-component decay for FAD in aqueous solution with pH-depending lifetimes and relative amplitudes. Circular dichroism data were analyzed by a multi-Gaussian fitting procedure and the trends of properly chosen parameters confirmed pH-depending changes. The comparison between the results obtained by these two optical techniques allowed us to improve the significance of the outcome of circular dichroism. This combined approach may provide a useful tool for biotechnological investigation.
ISSN:0175-7571
1432-1017
DOI:10.1007/s00249-019-01369-0