作者
Michael Nikolaev, E Round, I Gushchin, V Polovinkin, T Balandin, P Kuzmichev, V Shevchenko, V Borshchevskiy, A Kuklin, A Round, F Bernhard, D Willbold, G Büldt, V Gordeliy
发表日期
2017/3/1
期刊
Crystal Growth & Design
卷号
17
期号
3
页码范围
945-948
出版商
American Chemical Society
简介
Membrane-like nanodiscs (ND) have become an important tool for the cell-free expression, solubilization, folding, and in vitro structural and functional studies of membrane proteins (MPs). Direct crystallization of MPs embedded in NDs would be of high importance for structural biology. However, despite considerable efforts we have been as yet unable to obtain crystals suitable for X-ray crystallography. In the present work, we show that an ND-trapped MP can be transferred into the cubic phase and crystallized in meso. Bacteriorhodopsin (BR) reconstituted into nanodiscs was mixed with a lipidic mesophase and crystallization was induced by adding a precipitant. The resulting crystals diffract beyond 1.8 Å. The structure of BR was solved at 1.9 Å and found to be indistinguishable from previous structures obtained with the protein solubilized in detergent. We suggest the proposed protocol of in meso crystallization to …
引用总数
201720182019202020212022202351074787
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