5M7J

Blastochloris viridis photosynthetic reaction center structure using best crystal approach

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	FREE ELECTRON LASER
Source details	SLAC LCLS BEAMLINE CXI
Synchrotron site	SLAC LCLS
Beamline	CXI
Temperature [K]	293
Detector technology	PIXEL
Collection date	2011-02-19
Detector	CS-PAD CXI-1
Wavelength(s)	1.32
Spacegroup name	P 21 21 21
Unit cell lengths	57.900, 84.800, 384.300
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	52.761 - 3.500
R-factor	0.2542
Rwork	0.253
R-free	0.27310
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4cas
RMSD bond length	0.003
RMSD bond angle	0.701
Data reduction software	CrystFEL (0.6.2)
Refinement software	PHENIX ((1.10_2155: ???))

Data quality characteristics

	Overall
Low resolution limit [Å]	58.000
High resolution limit [Å]	3.500
Number of reflections	24815
<I/σ(I)>	6.1
Completeness [%]	99.4
Redundancy	59

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	BATCH MODE		293	Melted monoolein was thoroughly mixed in a ratio of 3:2 (v/v) with 0.1M HEPES, pH 7.5, 0.1 % LDAO until a viscous, transparent LCP was obtained. The formed phase was then transferred into a glass vial and sponge-phase-inducing solution (1:4 ratio) was added containing 16% Jeffamine M-600, 1M HEPES pH 7.9, 0.7 M Ammonium sulphate, 2.5% 1,2,3-Heptanetriol, which swell the cubic phase to sponge phase. After phase separation overnight, the upper phase (sponge phase) was harvested. Crystals were grown using batch crystallization in the lipidic-sponge phase. Equal amount of sponge phase and protein were mixed with 2/5 (v/v) of 1.2 M Tris-sodium citrate and allowed to incubate for several weeks.