5M7K

Blastochloris viridis photosynthetic reaction center - RC_vir_xfel

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	55.438 - 3.500
R-factor	0.254
Rwork	0.253
R-free	0.27380
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4cas
RMSD bond length	0.003
RMSD bond angle	0.712
Data scaling 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	25004
<I/σ(I)>	3.5
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.