Tug-of-war: molecular dynamometers against living cells for analyzing sub-piconewton interaction of a specific protein with the cell membrane,Chemical Science

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Tug-of-war: molecular dynamometers against living cells for analyzing sub-piconewton interaction of a specific protein with the cell membrane
Chemical Science ( IF 7.6 ) Pub Date : 2021-09-02 , DOI: 10.1039/d1sc03059k
Huipu Liu ₁ , Yunlong Chen ₁ , Jiawei Wang ₁ , Yuanjiao Yang ₁ , Huangxian Ju ₁

Affiliation

Protein–membrane interactions play important roles in signal transductions and functional regulation of membrane proteins. Here, we design a molecular dynamometer (MDM) for analyzing protein–membrane interaction on living cells. The MDM is constructed by assembling an artificial lipid bilayer and alkylated Cy3-DNA azide (azide-Cy3-Cx) on a silica bubble. After a functional aptamer is covalently anchored onto the corresponding target protein on a living cell through UV irradiation, azide-Cy3-Cx is conjugated with the aptamer through a click reaction to produce a “tug-of-war” between the MDM and the cell due to the buoyancy of the silica bubble. This induces the detachment of the protein from the cell membrane or the alkane terminal from the MDM enabling sub-piconewton embedding force measurement by changing the alkane chain length and simple fluorescence analysis. The successful analysis of embedding force variation of a protein on the cell membrane upon post-translational modifications demonstrates the practicability and expansibility of this method for mechanics-related research in biological systems.

中文翻译：

拔河比赛：针对活细胞的分子测力计，用于分析特定蛋白质与细胞膜的亚皮牛顿相互作用

蛋白质-膜相互作用在膜蛋白的信号转导和功能调节中起重要作用。在这里，我们设计了一个分子测力计 (MDM) 来分析活细胞上的蛋白质 - 膜相互作用。MDM 是通过在二氧化硅气泡上组装人工脂质双层和烷基化 Cy3-DNA 叠氮化物 (叠氮化物-Cy3-C x )来构建的。通过紫外线照射将功能性适体共价锚定到活细胞上的相应靶蛋白后，叠氮化物-Cy3-C x由于二氧化硅气泡的浮力，MDM 与细胞之间通过点击反应与适体结合，从而在 MDM 和细胞之间产生“拉锯战”。这会诱导蛋白质从细胞膜或烷烃末端与 MDM 分离，从而通过改变烷烃链长度和简单的荧光分析来实现亚皮克牛顿嵌入力的测量。翻译后修饰后蛋白质在细胞膜上的嵌入力变化的成功分析证明了该方法在生物系统力学相关研究中的实用性和可扩展性。

更新日期：2021-09-08

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