Effect of halogenated fluorescent compounds on bioluminescent reactions
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The paper investigates an application of luminescent bioassays to monitor the toxicity of organic halides. Effects of xanthene dyes (fluorescein, eosin Y, and erythrosin B), used as model compounds, on bioluminescent reactions of firefly Luciola mingrelica, marine bacteria Photobacterium leiognathi, and hydroid polyp Obelia longissima were studied. Dependence of bioluminescence quenching constants on the atomic weight of halogen substituents in dye molecules was demonstrated. Bacterial bioluminescence was shown to be most sensitive to heavy halogen atoms involved in molecular structure; hence, it is suitable for construction of sensors to monitor toxicity of halogenated compounds. Mechanisms of bioluminescence quenching—energy transfer processes, collisional interactions, and enzyme–dye binding—were considered. Changes of bioluminescence (BL) spectra in the presence of the dyes were analyzed. Interactions of the dyes with enzymes were studied using fluorescence characteristics of the dyes in steady-state and time-resolved experiments. The dependences of fluorescence anisotropy of enzyme-bound dyes, the average fluorescence lifetime, and the number of exponential components in fluorescence decay on the atomic weight of halogen substituents were demonstrated. The results are discussed in terms of “dark effect of heavy halogen atom” in the process of enzyme–dye binding; hydrophobic interactions were assumed to be responsible for the effect.
KeywordsBioluminescent assay Xanthene dyes Effect of heavy halogen atom Bioluminescent enzymes Fluorescence Lifetime Anisotropy
Adenosine-5-triphosphate disodium salt
β-Nicotinamide adenine dinucleotide reduced
The authors thank Professor L. Frank for obelin preparation and Professor A.G. Sizykh for discussion of the results. The work was supported by: the program “Molecular and Cellular Biology” of the RAS; the grant “Leading Scientific School” N 64987.2010.4, a Grant of the Ministry of Education and Science RF N 126.96.36.199/5309; the Federal Target Program “Research and scientific-pedagogical personnel of innovation in Russia”, 2009–2013, contract N 02.740.11.0766.
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