5NJ4

From macrocrystals to microcrystals: a strategy for membrane protein serial crystallography

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]	298
Detector technology	PIXEL
Collection date	2016-06-09
Detector	CS-PAD CXI-1
Wavelength(s)	1.307
Spacegroup name	P 43 21 2
Unit cell lengths	226.500, 226.500, 113.900
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	35.700 - 2.400
R-factor	0.16214
Rwork	0.161
R-free	0.18632
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2i5n
RMSD bond length	0.014
RMSD bond angle	1.960
Data reduction software	CrystFEL (0.6.2)
Data scaling software	CrystFEL (0.6.2)
Phasing software	PHASER
Refinement software	REFMAC (5.8.0158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	35.700	2.460
High resolution limit [Å]	2.400	2.400
Number of reflections	118454
<I/σ(I)>	11.2
Completeness [%]	100.0	100
Redundancy	1529	106
CC(1/2)	0.997	0.438

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.8	277	Macrocrystal growth: 10 mg/ml protein 3.6 M ammonium sulphate, 6 % w/v heptane-1,2,3-triol, 20 mM KH203/K2H03 pH 6.8, 0.1 % LDAO 20 ul sitting drop, 10 protein : 10 precipitant Microcrystals 8.5 mg/ml protein 18.5 ul sitting drop, 10 protein : 7.5 precipitant : 1 seed stock