TY  - JOUR
AU  - Nikolaev, Michael
AU  - Round, E.
AU  - Gushchin, Ivan
AU  - Polovinkin, V.
AU  - Balandin, Taras
AU  - Kuzmichev
AU  - Shevchenko, Vitaly
AU  - Borshchevskiy, Valentin
AU  - Kuklin, A.
AU  - Round, A.
AU  - Bernhard, F.
AU  - Willbold, Dieter
AU  - Büldt, Georg
AU  - Gordeliy, Valentin
TI  - Integral Membrane Proteins Can Be Crystallized Directly from Nanodiscs
JO  - Journal of crystal growth
VL  - 17
IS  - 3
SN  - 0022-0248
CY  - Amsterdam [u.a.]
PB  - Elsevier
M1  - FZJ-2017-03829
SP  - 945–948
PY  - 2017
AB  - 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 be generally applicable to ND-trapped MPs.
LB  - PUB:(DE-HGF)16
UR  - <Go to ISI:>//WOS:000395493900003
DO  - DOI:10.1021/acs.cgd.6b01631
UR  - https://juser.fz-juelich.de/record/830256
ER  -