6V91

Crystal structure of Stringent starvation protein A (BTH_I2974) from Burkholderia thailandensis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2019-11-21
Detector	RAYONIX MX-300
Wavelength(s)	0.97872
Spacegroup name	P 21 21 21
Unit cell lengths	60.380, 95.640, 108.850
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	33.920 - 1.900
R-factor	0.1756
Rwork	0.174
R-free	0.21490
Structure solution method	SAD
RMSD bond length	0.007
RMSD bond angle	0.852
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX (1.17.1)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.950
High resolution limit [Å]	1.900	8.500	1.900
Rmerge	0.047	0.027	0.520
Rmeas	0.051	0.031	0.570
Number of reflections	50160	614	3608
<I/σ(I)>	19.65	40.62	3.58
Completeness [%]	99.5	94.5	96.9
Redundancy	5.952	4.795	6.006
CC(1/2)	0.999	0.998	0.909

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		290	For native data collection: Anatrace Morpheus screen G7: 20mM of each Sodium formate, Ammonium acetate, Sodium citrate tribasic dihydrate, Sodium potassium tartrate tetrahydrate, Sodium oxamate: 100mM Na-HEPES / MOPS acid pH 7.5: 40% (V/V) Glycerol, 20% (w/V) PEG 4000: ButhA.17888.a.AE1 (labled as CrpaA.01056.a.AE1.PW38685) at 30.5mg/ml: 311984 g7: cryo: direct: puck ocs5-5. For phasing, a crystal from the same tray, condition H9 (20mM of each L-Na-Glutamate, Alanine (racemic), Glycine, Lysine HCl (racemic), Serine (racemic): 100mM Tris base / Bicine pH 8.5: 40% (v/v) PEG 500 MME, 20% (w/V) PEG 20000) was incubated for 30sec in a mix of 90% reservoir and 10% 5M NaI in ethylene glycol and vitrified without further cryoprotection. 360deg of data were collected in-house at Cu-Ka radiation.Tray: 311984 h9, puck ocs5-14.
1	VAPOR DIFFUSION, SITTING DROP		290	For native data collection: Anatrace Morpheus screen G7: 20mM of each Sodium formate, Ammonium acetate, Sodium citrate tribasic dihydrate, Sodium potassium tartrate tetrahydrate, Sodium oxamate: 100mM Na-HEPES / MOPS acid pH 7.5: 40% (V/V) Glycerol, 20% (w/V) PEG 4000: ButhA.17888.a.AE1 (labled as CrpaA.01056.a.AE1.PW38685) at 30.5mg/ml: 311984 g7: cryo: direct: puck ocs5-5. For phasing, a crystal from the same tray, condition H9 (20mM of each L-Na-Glutamate, Alanine (racemic), Glycine, Lysine HCl (racemic), Serine (racemic): 100mM Tris base / Bicine pH 8.5: 40% (v/v) PEG 500 MME, 20% (w/V) PEG 20000) was incubated for 30sec in a mix of 90% reservoir and 10% 5M NaI in ethylene glycol and vitrified without further cryoprotection. 360deg of data were collected in-house at Cu-Ka radiation.Tray: 311984 h9, puck ocs5-14.