5V33

R. sphaeroides photosythetic reaction center mutant - Residue L223, Ser to Trp - Room Temperature Structure Solved on X-ray Transparent Microfluidic Chip

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	298
Detector technology	PIXEL
Collection date	2016-08-04
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.000
Spacegroup name	P 42 21 2
Unit cell lengths	102.332, 102.332, 240.333
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	47.076 - 3.487
R-factor	0.2246
Rwork	0.221
R-free	0.26160
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4tqq
RMSD bond length	0.003
RMSD bond angle	0.702
Data reduction software	HKL-2000 (v704x)
Data scaling software	HKL-2000 (v704x)
Phasing software	PHASER
Refinement software	PHENIX

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	3.560
High resolution limit [Å]	3.500	9.480	3.500
Rmerge	0.290	0.148	0.720
Rmeas	0.322	0.172	0.793
Rpim	0.132	0.081	0.317
Total number of observations	84779
Number of reflections	15671
<I/σ(I)>	2.9
Completeness [%]	92.0	88.5	94.6
Redundancy	5.4	4.8	5.4
CC(1/2)		0.960	0.618

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	MICROFLUIDIC	7.8	293	Crystals were grown in microfluidic wells. LCPs were formulated by overlaying ~40 nl of protein solution over ~10 nl of solid monoolein. LCPs formed by diffusion during a 4 hour incubation. Precipitants were then introduced to the crystallization well, and crystals were observed after 3 days. Crystals were grown at 15-20 mg/mL protein concentration (in 10 mM Tris pH 7.8, 280 mM NaCl, 0.05% LDAO) with a precipitant of 1 M HEPES pH 7.5, 1.15 M (NH4)2(SO4) and 14-15 %w/v Jeffamine M-600.