通过控制连接体的内距实现基于能量转移的膜胆固醇识别

Yong Ho Cho, Tae Kyung Won, Dong June Ahn
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摘要

金纳米粒子(AuNPs)是能量转移系统中供体材料的良好候选材料,可通过 Au-S 键轻松地在其表面与各种配体功能化。环糊精(CD)因其独特的主客体相互作用结构而与荧光团形成包合物。在本研究中,我们利用不同长度的硫醇配体和识别胆固醇制备了 βCD 功能化 AuNPs,以证实基于能量转移的荧光开启机制。AuNP-βCD 与不同的硫醇配体共轭,淬灭荧光素(Fl)染料,形成 βCD-Fl 包合物。随着 AuNPs 与 βCD 之间距离的减小,淬灭效率也随之升高。由于胆固醇与 βCD 的结合亲和力更强,当胆固醇取代 Fl 时,淬灭的荧光得以恢复。胆固醇的识别效率也受到能量转移效应的影响,因为较短的βCD配体具有更高的荧光恢复率。此外,我们还制作了一种脂质体,在脂质双层膜中嵌入胆固醇,以模拟人血清中与脂质共存的胆固醇。这些细胞胆固醇加速了 Fl 分子的置换,从而使荧光回收率高于纯脂质。这些发现有望为使用 AuNPs 的胆固醇传感器或基于能量转移的生物传感器提供指导。
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Energy Transfer-Based Recognition of Membrane Cholesterol by Controlling Intradistance of Linker
Gold nanoparticles (AuNPs) are good candidates for donor material in energy transfer systems and can easily be functionalized with various ligands on the surface with Au–S bonding. Cyclodextrin (CD) forms inclusion complexes with fluorophores due to its unique structure for host–guest interaction. In this study, we fabricated βCD-functionalized AuNPs using different lengths of thiol ligands and recognized cholesterol to confirm the energy-transfer-based turn-on fluorescence mechanism. AuNP–βCD conjugated with various thiol ligands and quenched the fluorescein (Fl) dye, forming βCD-Fl inclusion complexes. As the distance between AuNPs and βCD decreased, the quenching efficiency became higher. The quenched fluorescence was recovered when the cholesterol replaced the Fl because of the stronger binding affinity of the cholesterol with βCD. The efficiency of cholesterol recognition was also affected by the energy transfer effect because the shorter βCD ligand had a higher fluorescence recovery. Furthermore, we fabricated a liposome with cholesterol embedded in the lipid bilayer membrane to mimic the cholesterol coexisting with lipids in human serum. These cellular cholesterols accelerated the replacement of the Fl molecules, resulting in a fluorescence recovery higher than that of pure lipid. These discoveries are expected to give guidance towards cholesterol sensors or energy-transfer-based biosensors using AuNPs.
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