2gnu
From Proteopedia
(New page: 200px<br /><applet load="2gnu" size="450" color="white" frame="true" align="right" spinBox="true" caption="2gnu, resolution 2.200Å" /> '''The crystallization...) |
|||
| Line 1: | Line 1: | ||
| - | [[Image:2gnu.gif|left|200px]]<br /><applet load="2gnu" size=" | + | [[Image:2gnu.gif|left|200px]]<br /><applet load="2gnu" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="2gnu, resolution 2.200Å" /> | caption="2gnu, resolution 2.200Å" /> | ||
'''The crystallization of reaction center from Rhodobacter sphaeroides occurs via a new route'''<br /> | '''The crystallization of reaction center from Rhodobacter sphaeroides occurs via a new route'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Bicontinuous lipidic cubic phases can be used as a host for growing | + | 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. |
==About this Structure== | ==About this Structure== | ||
| - | 2GNU is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Rhodobacter_sphaeroides Rhodobacter sphaeroides] with FE2, CL, BCL, BPH, U10, CDL and LDA as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http:// | + | 2GNU is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Rhodobacter_sphaeroides Rhodobacter sphaeroides] with <scene name='pdbligand=FE2:'>FE2</scene>, <scene name='pdbligand=CL:'>CL</scene>, <scene name='pdbligand=BCL:'>BCL</scene>, <scene name='pdbligand=BPH:'>BPH</scene>, <scene name='pdbligand=U10:'>U10</scene>, <scene name='pdbligand=CDL:'>CDL</scene> and <scene name='pdbligand=LDA:'>LDA</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2GNU OCA]. |
==Reference== | ==Reference== | ||
| Line 13: | Line 13: | ||
[[Category: Protein complex]] | [[Category: Protein complex]] | ||
[[Category: Rhodobacter sphaeroides]] | [[Category: Rhodobacter sphaeroides]] | ||
| - | [[Category: Cogdell, R | + | [[Category: Cogdell, R J.]] |
[[Category: Engstroem, S.]] | [[Category: Engstroem, S.]] | ||
| - | [[Category: Gardiner, A | + | [[Category: Gardiner, A T.]] |
[[Category: Katona, G.]] | [[Category: Katona, G.]] | ||
[[Category: Neutze, R.]] | [[Category: Neutze, R.]] | ||
[[Category: Snijder, A.]] | [[Category: Snijder, A.]] | ||
[[Category: Wadsten, P.]] | [[Category: Wadsten, P.]] | ||
| - | [[Category: Woehri, A | + | [[Category: Woehri, A B.]] |
[[Category: BCL]] | [[Category: BCL]] | ||
[[Category: BPH]] | [[Category: BPH]] | ||
| Line 31: | Line 31: | ||
[[Category: ubiquinone b]] | [[Category: ubiquinone b]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:33:37 2008'' |
Revision as of 15:33, 21 February 2008
|
The crystallization of reaction center from Rhodobacter sphaeroides occurs via a new route
Overview
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.
About this Structure
2GNU is a Protein complex structure of sequences from Rhodobacter sphaeroides with , , , , , and as ligands. Full crystallographic information is available from OCA.
Reference
Lipidic sponge phase crystallization of membrane proteins., Wadsten P, Wohri AB, Snijder A, Katona G, Gardiner AT, Cogdell RJ, Neutze R, Engstrom S, J Mol Biol. 2006 Nov 17;364(1):44-53. Epub 2006 Jul 7. PMID:17005199
Page seeded by OCA on Thu Feb 21 17:33:37 2008
Categories: Protein complex | Rhodobacter sphaeroides | Cogdell, R J. | Engstroem, S. | Gardiner, A T. | Katona, G. | Neutze, R. | Snijder, A. | Wadsten, P. | Woehri, A B. | BCL | BPH | CDL | CL | FE2 | LDA | U10 | Reaction centre | Ubiquinone b
