5V0S

Crystal structure of the ACT domain of prephenate dehydrogenase tyrA from Bacillus anthracis

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	2014-02-21
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.97872
Spacegroup name	P 65 2 2
Unit cell lengths	48.070, 48.070, 233.395
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	41.630 - 2.010
R-factor	0.1737
Rwork	0.171
R-free	0.23230
Structure solution method	SAD
RMSD bond length	0.011
RMSD bond angle	1.501
Data reduction software	HKL-3000
Data scaling software	HKL-2000
Phasing software	SHELX
Refinement software	REFMAC (5.8.0158)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.030
High resolution limit [Å]	2.000	5.430	2.000
Rmerge	0.053	0.037	0.619
Rmeas	0.057	0.040	0.653
Rpim	0.018	0.014	0.195
Number of reflections	11294		482
<I/σ(I)>	9.8		1.7
Completeness [%]	96.9	93.6	88.1
Redundancy	7.4	6.4	7.5
CC(1/2)		0.999	0.844

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	289	0.2 ul of 15 mg/ml protein in 20 mM HEPES pH 7.5, 500 mM NaCl, 5% Glycerol, and 10 mM BME were mixed with 0.2 ul of the MCSG Suite I condition #31 (0.1 M Tris pH=8.5, 0.2 M Calcium chloride, 25% w/v PEG 4000) and equilibrated against 1.5 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci). Before crystallization protein was incubated with 1/50 v/v of 2 mg/ml chymotrypsin solution at 289 K for 3 hours.