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Far-red pentamethine cyanine dyes as fluorescent probes for the detection of serum albumins

机译:远红五甲磺酰氨氨酸染料作为荧光探针检测血清白化素

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摘要

Benzothiazole based cyanine dyes with bridged groups in the pentamethine chain were studied as potential far-red fluorescent probes for protein detection. Spectral-luminescent properties were characterized for unbound dyes and in the presence of serum albumins (bovine (BSA), human (HSA), equine (ESA)), and globular proteins (β-lactoglobulin, ovalbumin). We have observed that the addition of albumins leads to a significant increase in dyes fluorescence intensity. However, the fluorescent response of dyes in the presence of other globular proteins was notably lower. The value of fluorescence quantum yield for dye bearing a sulfonate group complexed with HSA amounted to 42% compared with 0.2% for the free dye. The detection limit of HSA by this dye was greater than 0.004 mg ml−1 which indicates the high sensitivity of dye to low HSA concentrations. Modelling of structure of the dyes complexes with albumin molecules was performed by molecular docking. According to these data, dyes could bind to up to five sites on the HSA molecule; the most preferable are the haemin-binding site in subdomain IB and the dye-binding site in the pocket between subdomains IA, IIA and IIIA. This work confirms that pentamethine cyanine dyes could be proposed as powerful far-red fluorescent probes applicable for highly sensitive detection of albumins.
机译:基于苯并噻唑基吡啶染料与五甲基链条中的桥接基团进行了研究,作为蛋白质检测的潜在远红色荧光探针。光谱 - 发光性能的特征在于未结合的染料,并且在血清酶(牛(BSA),人(HSA),大床(ESA)和球状蛋白质(β-乳酰叶蛋白,卵巢)存在下。我们观察到,添加蛋白质导致染料荧光强度的显着增加。然而,在其他球状蛋白存在下染料的荧光响应尤其较低。含有HSA络合的磺酸盐基团的染料荧光量子产率的价值为42%,比自由染料为0.2%。该染料的HSA的检测极限大于0.004mg ML-1,表明染料对低HSA浓度的高灵敏度。用分子对接进行用白蛋白分子的染料复合物的结构建模。根据这些数据,染料可以在HSA分子上结合多达五个位点;最优选的是亚域IB的血液结合位点和亚域IA,IIA和IIIa之间的口袋中的染料结合位点。这项工作证实,五甲磺酸丁氰胺染料可以提出强大的远红荧光探针,适用于对白化素的高敏感性检测。

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