High‐Resolution AFM Imaging of Prestin Purified and Reconstituted into an Artificial Lipid Bilayer
High‐Resolution AFM Imaging of Prestin Purified and Reconstituted into an Artificial Lipid Bilayer
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Prestin 纯化并重构为人工脂质双层的高分辨率 AFM 成像
DOI：
10.1063/1.3658084
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
H. Hamana
中科院分区：
文献类型：
--
作者：
H. Wada;S. Kumano;M. Murakoshi;Koji Iida;H. Hamana
在本研究中，用原子力显微镜(AFM)观察纯化的Prestin。首先，通过沉积小的单层脂泡，在新裂解的云母盘上形成脂双层。这种双层膜通过与洗涤剂孵育而不稳定。然后，将纯化的Prestin添加到不稳定的脂双层中。最后，利用原子力显微镜获得了重组脂质双层的高分辨率图像。结果显示，Prestin包埋致密，直径为11.0±1.3 nm。从获得的图像中可以看到，Prestin的细胞质一侧形成了一个四峰一谷的环状结构。
In the present study, purified prestin was observed by atomic force microscopy (AFM). First, the lipid bilayer was formed on a freshly cleaved mica disk by the deposition of small unilamellar lipid vesicles. Such bilayer was destabilized by incubation with a detergent. Afterward, purified prestin was added to the destabilized lipid bilayer. Finally, a high resolution image of the prestin‐reconstituted lipid bilayer was acquired by AFM. As a result, densely embedded prestin with a diameter of 11.0±1.3 nm was visualized. From the obtained image, cytoplasmic side of prestin was found to form a ring‐like structure with four peaks and one valley at its center.