5VGM

Crystal structure of dihydroorotase pyrC from Vibrio cholerae in complex with zinc at 1.95 A resolution.

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	2017-03-12
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97872
Spacegroup name	P 1 21 1
Unit cell lengths	62.507, 57.257, 87.524
Unit cell angles	90.00, 105.09, 90.00

Refinement procedure

Resolution	20.000 - 1.950
R-factor	0.1695
Rwork	0.168
R-free	0.20300
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5V0G
RMSD bond length	0.010
RMSD bond angle	1.349
Data reduction software	HKL-3000
Data scaling software	SCALEPACK
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0158)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	20.000	20.000	1.980
High resolution limit [Å]	1.950	5.260	1.950
Rmerge	0.063	0.045	0.372
Rmeas	0.075	0.053	0.454
Rpim	0.040	0.028	0.257
Number of reflections	41257		1353
<I/σ(I)>	11.4		2.3
Completeness [%]	94.1	95.2	62.5
Redundancy	3.3	3.3	2.6
CC(1/2)	0.870	0.997	0.871

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	289	0.2 ul of 14 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide, 0.5 mM TCE, and 20 mM L-citruline were mixed with 0.2 ul of the Top96 screen condition #C11 (0.2 M Sodium Acetate 0.1 M Sodium Cacodylate: HCl, pH 6.5 30 % (w/v) PEG 8000), 0.1 ul of the Hampton Additive screen # 94 (7%v/v 1-Butanol) 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 1 hours.