6FV4

The structure of N-acetyl-D-glucosamine-6-phosphate deacetylase D267A mutant from Mycobacterium smegmatis in complex with N-acetyl-D-glucosamine-6-phosphate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I24
Synchrotron site	Diamond
Beamline	I24
Temperature [K]	100
Detector technology	PIXEL
Collection date	2017-07-14
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.968630
Spacegroup name	C 1 2 1
Unit cell lengths	164.766, 54.437, 110.262
Unit cell angles	90.00, 123.22, 90.00

Refinement procedure

Resolution	54.560 - 1.974
R-factor	0.1865
Rwork	0.184
R-free	0.23530
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6fv3
RMSD bond length	0.007
RMSD bond angle	0.901
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	PHENIX ((dev_2747: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	80.141	2.080
High resolution limit [Å]	1.974	1.974
Rmerge	0.128	0.822
Rmeas	0.152	0.987
Rpim	0.061	0.412
Number of reflections	55882	8181
<I/σ(I)>	9.1	2.7
Completeness [%]	96.8	98.1
Redundancy	6.5	6.3
CC(1/2)	0.993	0.841

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION		295.15	NagA crystals grew within a week at 295.15 K in 0.12 M monosaccharide mix (Morpheus, Molecular Dimensions), 0.1 M imidazole/MES pH 6.5, 20 % PEG 500 MME, 10 % w/v PEG 20000 with the addition of 10 mM CdCl2 additive. 5 mM GlcNAc6P was added and incubated at 277.14 K for 30 min before crystallization.