6BI4

2.9 Angstrom Resolution Crystal Structure of dTDP-Glucose 4,6-dehydratase (rfbB) from Bacillus anthracis str. Ames in Complex with NAD.

Replaces:  4EGB

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	2012-02-20
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97856
Spacegroup name	I 4 2 2
Unit cell lengths	165.557, 165.557, 292.121
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.950 - 2.910
R-factor	0.16928
Rwork	0.167
R-free	0.20436
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1r66
RMSD bond length	0.010
RMSD bond angle	1.667
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.950
High resolution limit [Å]	2.900	2.900
Rmerge	0.150	0.771
Rmeas	0.158	0.813
Rpim	0.051	0.256
Number of reflections	44858	2208
<I/σ(I)>	13	2.9
Completeness [%]	99.9	100
Redundancy	9.4	9.7
CC(1/2)		0.843

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	295	PROTEIN: TOTAL RFBB AND RFBC AT 7.5 MG/ML IN 10 MM TRIS HCL PH 8.3, 500 MM NACL, 5 MM BME, 10% GLYCEROL, 5 MM MGCL2, AND 1 MM NAD+. SCREEN: 0.1 M TRIS HCL PH 8.0, 1.56 M AMMONIUM SULFATE, AND 8.5% (W/V) PEG3350. CRYO-PROTECTANT SOLUTION: 1:1 RATIO OF 3.6 M AMMONIUM SULFATE AND 50% SUCROSE.