Integral Membrane Proteins Can Be Crystallized Directly from Nanodiscs | Zendy

Mikhail Nikolaev | Zendy; Ekaterina Round | Zendy; Ivan Gushchin | Zendy; Vitaly Polovinkin | Zendy; Taras Balandin | Zendy; Pavel Kuzmichev | Zendy; Vitaly Shevchenko | Zendy; Valentin Borshchevskiy | Zendy; A. I. Kuklin | Zendy; Adam Round | Zendy; Frank Bernhard | Zendy; Dieter Willbold | Zendy; Georg Büldt | Zendy; Valentin Gordeliy | Zendy

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Integral Membrane Proteins Can Be Crystallized Directly from Nanodiscs

Author(s) -

Mikhail Nikolaev,

Ekaterina Round,

Ivan Gushchin,

Vitaly Polovinkin,

Taras Balandin,

Pavel Kuzmichev,

Vitaly Shevchenko,

Valentin Borshchevskiy,

A. I. Kuklin,

Adam Round,

Frank Bernhard,

Dieter Willbold,

Georg Büldt,

Valentin Gordeliy

Publication year - 2017

Publication title -

crystal growth and design

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.966

H-Index - 155

eISSN - 1528-7505

pISSN - 1528-7483

DOI - 10.1021/acs.cgd.6b01631

Subject(s) - bacteriorhodopsin , crystallization , mesophase , crystallography , membrane protein , chemistry , integral membrane protein , membrane , folding (dsp implementation) , phase (matter) , solubilization , biophysics , biochemistry , organic chemistry , biology , engineering , electrical engineering

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

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