8QJD

T6SS-linked Rhs repeat protein - Salmonella bongori Rhs-core domain

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06SA
Synchrotron site	SLS
Beamline	X06SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-05-09
Detector	PSI PILATUS 6M
Wavelength(s)	1
Spacegroup name	P 1 21 1
Unit cell lengths	99.071, 118.260, 131.757
Unit cell angles	90.00, 105.52, 90.00

Refinement procedure

Resolution	49.170 - 2.203
R-factor	0.1837
Rwork	0.182
R-free	0.21100
Structure solution method	SIRAS
RMSD bond length	0.004
RMSD bond angle	0.666
Data reduction software	XDS
Data scaling software	XDS
Phasing software	BUCCANEER
Refinement software	PHENIX (1.20.1_4487+SVN)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.170	2.281
High resolution limit [Å]	2.203	2.203
Rmerge	0.080	0.855
Rmeas	0.087	0.929
Rpim	0.033
Number of reflections	147230	14230
<I/σ(I)>	17.62	2.32
Completeness [%]	99.6	97.11
Redundancy	6.84
CC(1/2)	0.999	0.836

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	292.15	Protein was diluted to 3 mg/ml using MonoQ buffer A and 1 ul was mixed with 2 ul of reservoir solution (0.1 M MES pH 7, 20% PEE 270, 3.67% acetone) in a hanging drop vapor diffusion experiment at 19 deg C to obtain the crystals for the native dataset used for SIRAS phasing and for refinement. Before the diffraction experiment, crystals were cryoprotected by soaking in a solution consisting of 0.1M MES pH7 and 35% PEE270. For the Hg(II) derivative dataset used in SIRAS phasing, the crystal was grown in a similar fashion but using a reservoir solution comprising 0.1 M HEPES pH 7, 20% PEE 270. Derivatisation was performed by soaking the crystal for a couple of minutes in reservoir solution supplemented with 12.5 mM thimerosal. Crystals were back soaked and cryoprotected in mother liquor with an increased PEE 270 concentration (35%).