Laboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave., 194064 Saint Petersburg, Russia.
Research Center for Molecular Mechanisms of Aging and Age-Related Diseases, Moscow Institute of Physics and Technology, 141700 Dolgoprudny, Russia.
Int J Mol Sci. 2021 Oct 15;22(20):11113. doi: 10.3390/ijms222011113.
The fluorescent dye BADAN (6-bromoacetyl-2-dimetylaminonaphtalene) is widely used in various fields of life sciences, however, the photophysical properties of BADAN are not fully understood. The study of the spectral properties of BADAN attached to a number of mutant forms of GGBP, as well as changes in its spectral characteristics during structural changes in proteins, allowed to shed light on the photophysical properties of BADAN. It was shown that spectral properties of BADAN are determined by at least one non-fluorescent and two fluorescent isomers with overlapping absorbing bands. It was found that BADAN fluorescence is determined by the unsolvated "PICT" (planar intramolecular charge transfer state) and solvated "TICT" (twisted intramolecular charge transfer state) excited states. While "TICT" state can be formed both as a result of the "PICT" state solvation and as a result of light absorption by the solvated ground state of the dye. BADAN fluorescence linked to GGBP/H152C apoform is quenched by Trp 183, but this effect is inhibited by glucose intercalation. New details of the changes in the spectral characteristics of BADAN during the unfolding of the protein apo and holoforms have been obtained.
荧光染料 BADAN（6-溴乙酰基-2-二甲氨基萘）广泛应用于生命科学的各个领域，然而，BADAN 的光物理性质尚未完全了解。研究 BADAN 与几种 GGBP 突变体形式的结合以及蛋白质结构变化过程中其光谱特性的变化，有助于阐明 BADAN 的光物理性质。结果表明，BADAN 的光谱性质至少由一个非荧光和两个荧光异构体决定，它们的吸收带重叠。研究发现，BADAN 的荧光由未溶剂化的“PICT”（平面分子内电荷转移态）和溶剂化的“TICT”（扭曲分子内电荷转移态）激发态决定。虽然“TICT”态既可以是“PICT”态溶剂化的结果，也可以是染料的溶剂化基态吸收光的结果。与 GGBP/H152C 去氧形式结合的 BADAN 荧光被色氨酸 183 猝灭，但这种效应被葡萄糖嵌入所抑制。在apo 和 holo 形式的蛋白质展开过程中，BADAN 光谱特性变化的新细节已经获得。
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