6BL6

Crystallization of lipid A transporter MsbA from Salmonella typhimurium

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL7-1
Synchrotron site	SSRL
Beamline	BL7-1
Temperature [K]	80
Detector technology	CCD
Collection date	2016-01-28
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.127085
Spacegroup name	P 21 21 2
Unit cell lengths	82.052, 154.819, 228.886
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.010 - 2.800
R-factor	0.27104
Rwork	0.270
R-free	0.30071
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3b5w
RMSD bond length	0.009
RMSD bond angle	1.368
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	REFMAC (5.8.0158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.010	2.850
High resolution limit [Å]	2.800	2.800
Rmerge	0.136	0.136
Rmeas	0.148
Rpim	0.057	0.057
Number of reflections	60504
<I/σ(I)>	10
Completeness [%]	84.9	82
Redundancy	5	4.1
CC(1/2)	0.893

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	293	MsbA purified in detergent micelles (UDM/FA231) is mixed with lipid A prior to crystallization and mixed 1: 1 with well solution 20 mM Tris-Hcl (pH 7.5) and 15-20 5 PEG 300 . Crystals appeared in month (approx.)