6U60

Crystal structure of prephenate dehydrogenase tyrA from Bacillus anthracis in complex with NAD and L-tyrosine

Replaces:  5USC

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2015-04-18
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97856
Spacegroup name	P 21 21 2
Unit cell lengths	106.850, 81.003, 83.597
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	36.480 - 2.100
R-factor	0.1634
Rwork	0.161
R-free	0.22030
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3ggg
RMSD bond length	0.008
RMSD bond angle	1.484
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0253)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.140
High resolution limit [Å]	2.100	5.700	2.100
Rmerge	0.075	0.034	1.302
Rmeas	0.080	0.037	1.411
Rpim	0.027	0.013	0.534
Number of reflections	42991	2322	2137
<I/σ(I)>	9.1		1.6
Completeness [%]	99.8	99.1	99.3
Redundancy	8.5	8.1	6.7
CC(1/2)	0.999	0.998	0.612

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	6.2	289	0.2 ul of 12 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 5% Glycerol, 10 mM BME, 5 mM NAD and 20 mM Tyrosine were mixed with 0.2 ul of the MCSG Suite 2 condition #81 (0.1 M Potassium/Sodium phosphate pH=6.2, 0.2M Sodium chloride, 20% w/v PEG 1000) and equilibrated against 1.5 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci). Before crystallization the protein was incubated with 1/50 v/v of 2 mg/ml chymotrypsin solution at 289 K for 3 hours