Lipidic sponge phase crystallization of membrane proteins.
Artikel i vetenskaplig tidskrift, 2006

Bicontinuous lipidic cubic phases can be used as a host for growing crystals of membrane proteins. Since the cubic phase is stiff, handling is difficult and time-consuming. Moreover, the conventional cubic phase may interfere with the hydrophilic domains of membrane proteins due to the limited size of the aqueous pores. Here, we introduce a new crystallization method that makes use of a liquid analogue of the cubic phase, the sponge phase. This phase facilitates a considerable increase in the allowed size of aqueous domains of membrane proteins, and is easily generalised to a conventional vapour diffusion crystallisation experiment, including the use of nanoliter drop crystallization robots. The appearance of the sponge phase was confirmed by visual inspection, small-angle X-ray scattering and NMR spectroscopy. Crystals of the reaction centre from Rhodobacter sphaeroides were obtained by a conventional hanging-drop experiment, were harvested directly without the addition of lipase or cryoprotectant, and the structure was refined to 2.2 Angstroms resolution. In contrast to our earlier lipidic cubic phase reaction centre structure, the mobile ubiquinone could be built and refined. The practical advantages of the sponge phase make it a potent tool for crystallization of membrane proteins.

Lipids

chemistry

X-Ray Diffraction

Protein Conformation

Binding Sites

Crystallization

chemistry

Rhodobacter sphaeroides

Photosynthetic Reaction Center Complex Proteins

Molecular Structure

Membrane Proteins

chemistry

chemistry

Författare

Pia Jeanette Hindrichsen

Chalmers, Kemi- och bioteknik, Teknisk ytkemi

Annemarie Wöhri

Chalmers, Kemi- och bioteknik, Molekylär bioteknik

Arjan Snijder

Chalmers, Kemi- och bioteknik, Molekylär bioteknik

Gergely Katona

Göteborgs universitet

Alastair T Gardiner

University of Glasgow

Richard J Cogdell

University of Glasgow

Richard Neutze

Göteborgs universitet

Sven Engström

Chalmers, Kemi- och bioteknik, Teknisk ytkemi

Journal of Molecular Biology

0022-2836 (ISSN)

Vol. 364 1 44-53

Ämneskategorier

Strukturbiologi

DOI

10.1016/j.jmb.2006.06.043

PubMed

17005199