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基于 QCM-D 的研究：脂质膜表面和 MUA 单层上 DNA 适体与细胞色素 C 相互作用分析。

Analysis of the Interaction between DNA Aptamers and Cytochrome C on the Surface of Lipid Films and on the MUA Monolayer: A QCM-D Study.

机构信息

Faculty of Mathematics, Physics and Informatics, Comenius University, Mlynská dolina F1, 842 48 Bratislava, Slovakia.

出版信息

Biosensors (Basel). 2023 Feb 9;13(2):251. doi: 10.3390/bios13020251.

DOI:10.3390/bios13020251
PMID:36832017
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9953847/
Abstract

We analyzed the possibility of the detection of cytochrome c (cyt c) being physically adsorbed on lipid films or covalently bounded to 11-mercapto-1-undecanoic acid (MUA) chemisorbed on the gold layer using quartz crystal microbalance with dissipation monitoring (QCM-D). The negatively charged lipid film composed of a mixture of zwitterionic DMPC and negatively charged DMPG phospholipids at a molar ratio of 1:1 allowed the formation of a stable cyt c layer. Addition of DNA aptamers specific to cyt c, however, resulted in removal of cyt c from the surface. The interaction of cyt c with the lipid film and its removal by DNA aptamers were accompanied by changes in viscoelastic properties evaluated using the Kelvin-Voigt model. Cyt c covalently bound to MUA also provided a stable protein layer already at its relatively low concentrations (0.5 μM). A decrease in the resonant frequency following the addition of gold nanowires (AuNWs) modified by DNA aptamers was observed. The interaction of aptamers with cyt c on the surface can be a combination of specific and non-specific interactions due to electrostatic forces between negatively charged DNA aptamers and positively charged cyt c.

摘要

我们使用石英晶体微天平（QCM-D）研究了细胞色素 c（cyt c）物理吸附在脂质膜上或与通过金层化学吸附的 11-巯基-1-十一酸（MUA）共价结合的可能性。由两性离子 DMPC 和带负电荷的 DMPG 磷脂以 1:1 的摩尔比组成的带负电荷的脂质膜允许形成稳定的 cyt c 层。然而，添加针对 cyt c 的 DNA 适体导致 cyt c 从表面去除。cyt c 与脂质膜的相互作用及其被 DNA 适体去除伴随着使用 Kelvin-Voigt 模型评估的粘弹性性质的变化。即使在相对较低的浓度（0.5 μM）下，共价结合到 MUA 的 cyt c 也能提供稳定的蛋白质层。添加经 DNA 适体修饰的金纳米线（AuNWs）后，观察到共振频率降低。由于带负电荷的 DNA 适体和带正电荷的 cyt c 之间的静电力，适体与表面上的 cyt c 的相互作用可以是特异性和非特异性相互作用的组合。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/32de43d48220/biosensors-13-00251-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/21299e7a4f34/biosensors-13-00251-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/c82fcc3fb449/biosensors-13-00251-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/3bc151bf4015/biosensors-13-00251-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/02de414384bc/biosensors-13-00251-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/6cad70ab598c/biosensors-13-00251-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/32de43d48220/biosensors-13-00251-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/21299e7a4f34/biosensors-13-00251-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/c82fcc3fb449/biosensors-13-00251-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/3bc151bf4015/biosensors-13-00251-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/02de414384bc/biosensors-13-00251-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/6cad70ab598c/biosensors-13-00251-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c1a9/9953847/32de43d48220/biosensors-13-00251-g006.jpg

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