High-Resolution Structure of a Membrane Protein Transferred from Amphipol to a Lipidic Mesophase

Amphipols (APols) have become important tools for the stabilization, folding, and in vitro structural and functional studies of membrane proteins (MPs). Direct crystallization of MPs solubilized in APols would be of high importance for structural biology. However, despite considerable efforts, it is...

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Published inThe Journal of membrane biology Vol. 247; no. 9-10; pp. 997 - 1004
Main Authors Polovinkin, V., Gushchin, I., Sintsov, M., Round, E., Balandin, T., Chervakov, P., Schevchenko, V., Utrobin, P., Popov, A., Borshchevskiy, V., Mishin, A., Kuklin, A., Willbold, D., Chupin, V., Popot, J.-L., Gordeliy, V.
Format Journal Article
LanguageEnglish
Published New York Springer US 01.10.2014
Springer Nature B.V
Springer Verlag
Subjects
Bacteriorhodopsins - chemistry
Bacteriorhodopsins - ultrastructure
Biochemistry
Biochemistry, Molecular Biology
Crystallization - methods
Human Physiology
Hydrophobic and Hydrophilic Interactions
Life Sciences
Lipid Bilayers - chemistry
Lipids
Membrane Proteins - chemistry
Membrane Proteins - ultrastructure
Membranes
Polymers
Polymers - chemistry
Propylamines - chemistry
Protein Conformation
Proteins
Solubility
Solutions
Structural Biology
Surface-Active Agents - chemistry
Bacteriorhodopsin
In meso crystallization
Monoolein
Amphipol
Membrane protein crystallization
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Summary:Amphipols (APols) have become important tools for the stabilization, folding, and in vitro structural and functional studies of membrane proteins (MPs). Direct crystallization of MPs solubilized in APols would be of high importance for structural biology. However, despite considerable efforts, it is still not clear whether MP/APol complexes can form well-ordered crystals suitable for X-ray crystallography. In the present work, we show that an APol-trapped MP can be crystallized in meso. Bacteriorhodopsin (BR) trapped by APol A8-35 was mixed with a lipidic mesophase, and crystallization was induced by adding a precipitant. The crystals diffract beyond 2 Å. The structure of BR was solved to 2 Å and found to be indistinguishable from previous structures obtained after transfer from detergent solutions. We suggest the proposed protocol of in meso crystallization to be generally applicable to APol-trapped MPs.
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ISSN:0022-2631
1432-1424
DOI:10.1007/s00232-014-9700-x