Enhanced EGFP fluorescence emission in presence of PEG aqueous solutions and PIB1000-PEG6000-PIB1000 copolymer vesicles.
Enhanced EGFP fluorescence emission in presence of PEG aqueous solutions and PIB1000-PEG6000-PIB1000 copolymer vesicles.
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DOI：
10.1155/2013/329087
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发表时间：
2013
影响因子：
--
通讯作者：
Fioroni M
中科院分区：
文献类型：
--
作者：
Muhammad N;Kryuchkova N;Dworeck T;Rodríguez-Ropero F;Fioroni M
与 PIB1000-PEG6000-PIB1000 囊泡表面相互作用的 EGFP 构建体报告了约 2 倍的荧光发射增强。由于将两亲性肽插入 PIB 核心的结构性质，EGFP 有望经历“纯”PEG 环境。为了解释这一现象，使用了不同分子量和浓度的 PEG/水溶液。在蛋白质/PEG 摩尔比约为 1:10 时，观察到荧光发射的增加达到与 PEG 分子量相关的稳定状态。在甘油水溶液存在下的平行实验确实显示出轻微的荧光增强，但是从更高的浓度开始。对纯水、甘油和 PEG 水溶液中的 EGFP 进行分子动力学模拟，结果表明 PEG 分子倾向于“包裹”蛋白质，从而形成局部 PEG 浓度高于其本体浓度的微环境。由于荧光发射可能受到蛋白质周围折射率的干扰，因此 PEG 分子的聚集在极低浓度下即可诱导增强的荧光发射。当与分子生物学实验中报道的 EGFP 的使用相关时，这些发现可能很重要。
An EGFP construct interacting with the PIB1000-PEG6000-PIB1000 vesicles surface reported a ~2-fold fluorescence emission enhancement. Because of the constructs nature with the amphiphilic peptide inserted into the PIB core, EGFP is expected to experience a “pure” PEG environment. To unravel this phenomenon PEG/water solutions at different molecular weights and concentrations were used. Already at ~1 : 10 protein/PEG molar ratio the increase in fluorescence emission is observed reaching a plateau correlating with the PEG molecular weight. Parallel experiments in presence of glycerol aqueous solutions did show a slight fluorescence enhancement however starting at much higher concentrations. Molecular dynamics simulations of EGFP in neat water, glycerol, and PEG aqueous solutions were performed showing that PEG molecules tend to “wrap” the protein creating a microenvironment where the local PEG concentration is higher compared to its bulk concentration. Because the fluorescent emission can be perturbed by the refractive index surrounding the protein, the clustering of PEG molecules induces an enhanced fluorescence emission already at extremely low concentrations. These findings can be important when related to the use of EGFP as reported in molecular biology experiments.
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