Rapid preparation of nanodiscs for biophysical studies.
Rapid preparation of nanodiscs for biophysical studies.
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DOI：
10.1016/j.abb.2021.109051
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发表时间：
2021-11-15
影响因子：
3.9
通讯作者：
Glover KJ
中科院分区：
文献类型：
--
作者：
Julien JA;Fernandez MG;Brandmier KM;Del Mundo JT;Bator CM;Loftus LA;Gomez EW;Gomez ED;Glover KJ
纳米圆盘是一种圆盘状的实体，含有一个由两亲性螺旋环包裹的中心脂质双层，已成为一种领先的天然样膜模拟物。目前用于形成纳米盘的方法涉及产生含有膜支架蛋白质、脂质和去污剂的混合胶束溶液，随后是透析和/或与吸附珠孵育的耗时过程（3-12小时）以从样品中除去去污剂分子。相比之下，本文所述的方法提供了一种使用Sephadex® G-25树脂从样品中除去去污剂在几分钟内（<15分钟）制备纳米盘的简便且快速的程序。采用一系列生物物理技术，包括分析超速离心、动态光散射、凝胶过滤色谱、圆二色性光谱和低温电子显微镜来明确证实通过这种方法形成的聚集体确实是纳米盘。我们相信，这种方法将是有吸引力的时间敏感和高通量的实验。
Nanodiscs, which are disc-shaped entities that contain a central lipid bilayer encased by an annulus of amphipathic helices, have emerged as a leading native-like membrane mimic. The current approach for the formation of nanodiscs involves the creation of a mixed-micellar solution containing membrane scaffold protein, lipid, and detergent followed by a time consuming process (3–12 h) of dialysis and/or incubation with sorptive beads to remove the detergent molecules from the sample. In contrast, the methodology described herein provides a facile and rapid procedure for the preparation of nanodiscs in a matter of minutes (<15 min) using Sephadex® G-25 resin to remove the detergent from the sample. A panoply of biophysical techniques including analytical ultracentrifugation, dynamic light scattering, gel filtration chromatography, circular dichroism spectroscopy, and cryogenic electron microscopy were employed to unequivocally confirm that aggregates formed by this method are indeed nanodiscs. We believe that this method will be attractive for time-sensitive and high-throughput experiments.
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影响因子：
7.7
作者：
Matthies D;Bae C;Toombes GE;Fox T;Bartesaghi A;Subramaniam S;Swartz KJ
通讯作者：
Swartz KJ
影响因子：
2.9
作者：
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通讯作者：
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影响因子：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
3
作者：
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通讯作者：
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