Detecting Single-Molecule Dynamics on Lipid Membranes with Quenchers-in-a-Liposome FRET
Detecting Single-Molecule Dynamics on Lipid Membranes with Quenchers-in-a-Liposome FRET
复制标题
使用脂质体淬灭剂 FRET 检测脂膜上的单分子动力学
DOI：
10.1002/anie.201813888
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Lu Ying
中科院分区：
文献类型：
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作者：
Ma Dong-Fei;Xu Chun-Hua;Hou Wen-Qing;Zhao Chun-Yu;Ma Jian-Bing;Huang Xing-Yuan;Jia Qi;Ma Lu;Diao Jiajie;Liu Cong;Li Ming;Lu Ying
跟踪膜相互作用分子并可视化其构象动力学是理解其功能的关键。然而，精确探测分子相对于膜的位置是具有挑战性的。在本文中，报道了一种称为LipoFRET的单分子方法来评估分子和脂质体之间的相互作用。它利用FRET在连接到生物分子的单个荧光团和脂质体中的许多猝灭剂之间的优势。该方法用于表征α-突触核蛋白（α-syn）与膜之间的相互作用。这些结果表明，α-syn的N-末端插入膜中，并在不同深度之间自发地转变。相比之下，α-syn的C-末端尾部受钙离子调节，并以两种构象漂浮在溶液中。LipoFRET是一种强大的工具，可以在单分子水平上以亚纳米精度研究膜相互作用的生物分子。
Tracking membrane‐interacting molecules and visualizing their conformational dynamics are key to understanding their functions. It is, however, challenging to accurately probe the positions of a molecule relative to a membrane. Herein, a single‐molecule method, termed LipoFRET, is reported to assess interplay between molecules and liposomes. It takes advantage of FRET between a single fluorophore attached to a biomolecule and many quenchers in a liposome. This method was used to characterize interactions between α‐synuclein (α‐syn) and membranes. These results revealed that the N‐terminus of α‐syn inserts into the membrane and spontaneously transitions between different depths. In contrast, the C‐terminal tail of α‐syn is regulated by calcium ions and floats in solution in two conformations. LipoFRET is a powerful tool to investigate membrane‐interacting biomolecules with sub‐nanometer precision at the single‐molecule level.