Watching Three-Dimensional Movements of Single Membrane Proteins in Lipid Bilayers
Watching Three-Dimensional Movements of Single Membrane Proteins in Lipid Bilayers
复制标题
观察脂质双层中单膜蛋白的三维运动
DOI：
10.1021/acs.biochem.8b00253
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发表时间：
2018
期刊：
影响因子：
2.9
通讯作者：
Li Ming
中科院分区：
文献类型：
--
作者：
Ma Li;Li Ying;Ma Jianbing;Hu Shuxin;Li Ming
由于缺乏合适的工具来检测单个蛋白在生物膜的0.4nm范围内的位置变化，因此评估蛋白质-膜相互作用具有挑战性。我们最近开发了一种测定，称为表面诱导荧光衰减（SIFA）。它能够跟踪支持的脂质双层中的单标记膜蛋白的垂直和横向动态运动。与FRET（荧光共振能量转移）原理类似，SIFA利用荧光团到吸光表面的能量转移来确定距离表面2-8 nm处的距离。通过用适当的荧光团标记蛋白质并使用氧化石墨烯作为二维猝灭剂，我们表明SIFA能够监测荧光团标记的蛋白质的三维运动，不仅在氧化石墨烯顶部的脂质双层内部，而且在脂质双层上方。我们的数据表明，SIFA是一种非常适合的方法来研究蛋白质和膜之间的相互作用。
It is challenging to assess protein–membrane interactions because of the lack of appropriate tools to detect position changes of single proteins in the ∼4 nm range of biological membranes. We developed an assay recently, termed surface-induced fluorescence attenuation (SIFA). It is able to track both vertical and lateral dynamic motion of singly labeled membrane proteins in supported lipid bilayers. Similar to the FRET (fluorescence resonance energy transfer) principle, SIFA takes advantage of the energy transfer from a fluorophore to a light-absorbing surface to determine the distance at 2–8 nm away from the surface. By labeling a protein with a proper fluorophore and using graphene oxide as a two-dimensional quencher, we showed that SIFA is capable of monitoring three-dimensional movements of the fluorophore-labeled protein not only inside but also above the lipid bilayer atop the graphene oxide. Our data show that SIFA is a well-suited method to study the interplay between proteins and membranes.