Mechanism of Budded Virus Envelope Fusion into a Planar Bilayer Lipid Membrane on a SiO2 Substrate
Mechanism of Budded Virus Envelope Fusion into a Planar Bilayer Lipid Membrane on a SiO2 Substrate
复制标题
DOI：
10.1021/acs.langmuir.1c03456
复制
发表时间：
2022-04-18
期刊：
影响因子：
3.9
通讯作者：
Sumitomo, Koji
中科院分区：
文献类型：
--
作者：
Oshima, Azusa;Nakanishi, Kohei;Sumitomo, Koji
人工平面双层类脂膜（BLM）是在物理和化学控制条件下的细胞系统的简单模型，并且它们已被用于研究膜蛋白活性。杆状病毒-出芽病毒（BV）系统可以表达重组膜蛋白。在这项研究中，针对膜蛋白重建，我们研究了融合的BV含有重组膜蛋白到人工平面BLMs上的Si微孔基板。BV与BLM的融合取决于溶液的pH值，在较低的pH值下，BV与BLM的融合增强。基于光漂白后荧光恢复（FRAP）测量，评估了BV的融合状态，并确认在低pH值下完全融合。在BLM的独立部分和支撑部分也观察到膜蛋白的荧光标记。这些结果表明，BLM作为一个平台，检查详细的融合动力学的BV的有效性。此外，这项研究表明，BV的融合是一种很有前途的方法，用于重建膜蛋白的人工独立的BLMs的生物器件的发展，我们可以检查膜蛋白的活性。
Artificial planar bilayer lipid membranes (BLMs) are simple models of cellular systems under physically and chemically controlled conditions, and they have been used to investigate membrane protein activity. Baculovirus-budded virus (BV) systems can express recombinant membrane proteins. In this study, aiming for membrane protein reconstitution, we examined the fusion of BVs containing recombinant membrane proteins into artificial planar BLMs on a Si microwell substrate. BV fusion with the BLMs depended on the pH of the solution, and it was enhanced at lower pH. Based on fluorescence recovery after photobleaching (FRAP) measurement, the fusion state of BVs was evaluated, and full fusion at low pH was confirmed. The fluorescent labeling the membrane proteins was also observed in the freestanding part of the BLMs as well as in the supported part. These results demonstrate the effectiveness of BLMs as a platform to examine detailed fusion dynamics of BVs. Furthermore, this study revealed that the fusion of BVs is a promising method for reconstituting membrane proteins to artificial freestanding BLMs for the development of biodevices with which we can examine membrane protein activity.