Effects of alcohols on fluorescence intensity and color of a discharged-obelin-based biomarker

Тип публикации: статья из журнала

Год издания: 2014

Идентификатор DOI: 10.1007/s00216-014-7685-z

Ключевые слова: Fluorescent protein, Biomarker, Photoprotein obelin, Red emission, Alcohols, Proton transfer, Amino acids, Bioluminescence, Biomarkers, Calcium, Color, Contrast media, Emission spectroscopy, Sugar substitutes, Excitation wavelength, Fluorescence intensities, Fluorescent intensity, Inhibition coefficient, Light induced fluorescence, Photoproteins, Red emissions, Fluorescence, Coelenterata, alcohol derivative, biological marker, obelin, photoprotein, animal, chemistry, Hydrozoa, spectrofluorometry, Animals, Biological Markers, Luminescent Proteins, Spectrometry, Fluorescence

Аннотация: Photoproteins are responsible for bioluminescence of marine coelenterates; bioluminescent and fluorescent biomarkers based on photoproteins are useful for monitoring of calcium-dependent processes in medical investigations. Here, we present the analysis of intensity and color of light-induced fluorescence of Ca2+-discharged photoprotein obelin in the presence of alcohols (ethanol and glycerol). Complex obelin spectra obtained at different concentrations of the alcohols at 350- and 280-nm excitation (corresponding to polypeptide-bound coelenteramide and tryptophan absorption regions) were deconvoluted into Gaussian components; fluorescent intensity and contributions of the components to experimental spectra were analyzed. Five Gaussian components were found in different spectral regions-ultraviolet (tryptophan emission), blue-green (coelenteramide emission), and red (hypothetical indole-coelenteramide exciplex emission). Inhibition coefficients and contributions of the components to experimental fluorescent spectra showed that presence of alcohols increased contributions of ultraviolet, violet, and red components, but decreased contributions of components in the blue-green region. The effects were related to (1) changes of proton transfer efficiency in fluorescent S*(1) state of coelenteramide in the obelin active center and (2) formation of indole-coelenteramide exciplex at 280-nm photoexcitation. The data show that variation of fluorescence color and intensity in the presence of alcohols and dependence of emission spectra on excitation wavelength should be considered while applying the discharged obelin as a fluorescence biomarker.

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Издание

Журнал: ANALYTICAL AND BIOANALYTICAL CHEMISTRY

Выпуск журнала: Vol. 406, Is. 12

Номера страниц: 2965-2974

ISSN журнала: 16182642

Место издания: HEIDELBERG

Издатель: SPRINGER HEIDELBERG

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