Lipidic sponge phase crystallization of membrane proteins.

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.

Wöhri, Annemarie,1976Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)wohri (författare)
Snijder, Arjan,1971Chalmers tekniska högskola,Chalmers University of Technology (författare)
Katona, Gergely,1975Gothenburg University,Göteborgs universitet,Institutionen för kemi,Department of Chemistry,University of Gothenburg(Swepub:gu)xkatge (författare)
Gardiner, Alastair TUniversity of Glasgow (författare)
Cogdell, Richard JUniversity of Glasgow (författare)
Neutze, Richard,1969Gothenburg University,Göteborgs universitet,Institutionen för kemi,Department of Chemistry,University of Gothenburg(Swepub:cth)xneuri (författare)
Engström, Sven,1951Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)sv (författare)
Hindrichsen, Pia Jeanette,1969Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)wadsten (författare)
Chalmers tekniska högskolaInstitutionen för kemi (creator_code:org_t)

Ingår i:Journal of molecular biology: Elsevier BV364:1, s. 44-530022-28361089-8638

Av författaren/redakt...: Wadsten, Pia, 19 ...; Wöhri, Annemarie ...; Snijder, Arjan, ...; Katona, Gergely, ...; Gardiner, Alasta ...; Cogdell, Richard ...; visa fler...; Neutze, Richard, ...; Engström, Sven, ...; Hindrichsen, Pia ...; visa färre...

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