N.M. Emanuel Institute of Biochemical Physics, Russian Academy of Sciences, Kosygin St. 4, Moscow 119334, Russia.
N.M. Emanuel Institute of Biochemical Physics, Russian Academy of Sciences, Kosygin St. 4, Moscow 119334, Russia.
Spectrochim Acta A Mol Biomol Spectrosc. 2017 Apr 15;177:93-96. doi: 10.1016/j.saa.2017.01.033. Epub 2017 Jan 18.
The noncovalent interaction of the polymethine dye probe 3,3',9-trimethylthiacarbocyanine iodide (Cyan 2) with chondroitin-4-sulfate (C4S) in buffer solutions with different pH and in water in the absence of buffers has been studied by spectral-fluorescent methods. It has been shown that in all media studied, at relatively high concentrations, the dye is bound to C4S mainly as a monomer, which is accompanied by a steep rise of fluorescence (the intermediate formation of dye aggregates on the biopolymer is also observed). From the dependence of the fluorescence quantum yield on the concentration of C4S, the parameters of binding of the dye monomer to C4S were obtained: the effective binding constant K, the number of the monomeric C4S units n per one dye monomer bound to C4S, and the fluorescence quantum yield of the bound dye monomer Φ. The dependence of Φ (and K) on рН of the medium is not monotonic: it has a minimum in the region of neutral pH and a growth in the regions of acid and basic pH. This can be explained by changing the charge of a C4S macromolecule as a function of pH and related conformational alterations in the biopolymer, which can affect the rigidity of a dye molecule and the energy of its interaction with the biopolymer.
本文采用光谱荧光法研究了聚甲川染料探针 3,3',9-三甲基噻嗪二碘化物(Cyan 2)与软骨素 4-硫酸盐(C4S)在不同 pH 值的缓冲溶液中和无缓冲剂的水中的非共价相互作用。结果表明,在所有研究的介质中,在相对较高的浓度下,染料主要以单体形式与 C4S 结合,这伴随着荧光的急剧上升(也观察到在生物聚合物上染料聚集体的中间形成)。从 C4S 浓度对染料单体荧光量子产率的依赖关系中,得到了染料单体与 C4S 结合的参数:有效结合常数 K、与一个 C4S 单体结合的单体 C4S 单元 n 的数量以及结合染料单体的荧光量子产率 Φ。Φ(和 K)对介质 pH 值的依赖性不是单调的:在中性 pH 区域有一个最小值,在酸性和碱性 pH 区域有一个增长。这可以用 C4S 大分子的电荷随 pH 值的变化以及生物聚合物中相关的构象改变来解释,这可能影响染料分子的刚性及其与生物聚合物的相互作用能。