PDBsum entry 2gnu

Photosynthesis

PDB id: 2gnu

Contents

Protein chains

235 a.a. *

281 a.a. *

300 a.a. *

Ligands

LDA

BCL ×4

U10 ×2

BPH ×2

CDL

Metals

FE2

_CL

Waters ×155

* Residue conservation analysis

PDB id:

2gnu

Name:

Photosynthesis

Title:

The crystallization of reaction center from rhodobacter sphaeroides occurs via a new route

Structure:

Reaction center protein h chain. Chain: h. Fragment: reaction center protein h chain, cytoplasmic domain, residue 11-245. Synonym: photosynthetic reaction center h subunit. Reaction center protein l chain. Chain: l. Synonym: photosynthetic reaction center l subunit. Reaction center protein m chain.

Source:

Rhodobacter sphaeroides. Organism_taxid: 1063. Strain: r26. Other_details: lacking carotenoids. Other_details: lacking carotenoids

Biol. unit:

Trimer (from PQS)

Resolution:

2.20Å R-factor: 0.200 R-free: 0.246

Authors:

P.Wadsten,A.B.Woehri,A.Snijder,G.Katona,A.T.Gardiner,R.J.Cogdell, R.Neutze,S.Engstroem

Key ref:

P.Wadsten et al. (2006). Lipidic sponge phase crystallization of membrane proteins. J Mol Biol, 364, 44-53. PubMed id: 17005199 DOI: 10.1016/j.jmb.2006.06.043

Date:

11-Apr-06 Release date: 07-Nov-06

Protein chain

Q7B302  (Q7B302_CERSP) -  Photosynthetic reaction center subunit H from Cereibacter sphaeroides

Seq: 260 a.a.

Struc: 235 a.a.

Protein chain

P0C0Y8  (RCEL_CERSP) -  Reaction center protein L chain from Cereibacter sphaeroides

Seq: 282 a.a.

Struc: 281 a.a.

Protein chain

P0C0Y9  (RCEM_CERSP) -  Reaction center protein M chain from Cereibacter sphaeroides

Seq: 308 a.a.

Struc: 300 a.a.

DOI no: 10.1016/j.jmb.2006.06.043

J Mol Biol 364:44-53 (2006)

PubMed id: 17005199

Lipidic sponge phase crystallization of membrane proteins.

P.Wadsten, A.B.Wöhri, A.Snijder, G.Katona, A.T.Gardiner, R.J.Cogdell, R.Neutze, S.Engström.

ABSTRACT

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.

Selected figure(s)

Figure 2.
Figure 2. Crystals grown directly from the L[3] phase using MO and buffer solution. (a) RC crystal fished directly from the L[3] phase. (b) and (c) Crystals of RC grown in 20% jeffamine M600 (pH 8.1).

Figure 4.
Figure 4. Q[B] binding sites of RCsph. All pictures are represented in stereoview. (a) Arrangement of the cofactors Q[A] and Q[B], subunits H, L, M and the location of the membrane in RCsph. The colouring scheme is: H, L, M subunit (dark grey), cofactors Q[A] (red), Q[B] (orange). The approximate position of the membrane is indicated by the light grey square. (b) Refined electron density for the Q[B] binding pocket. The 2F[o] – F[c] map is contoured at 1σ. (c) Simulated-annealing omit map omitting Q[B] contoured at 1σ. (d) Superposition of ubiquinone in the Q[B] binding pocket. Colour code and PDB entries: red, 1YST;^43 light green, 1AIG;^45 dark green, 1AIJ;^45 blue, 1RG5;^44 orange, present structure 2GNU.

The above figures are reprinted by permission from Elsevier: J Mol Biol (2006, 364, 44-53) copyright 2006.

Figures were selected by the author.