5F23

Crystal structure of NH(3)-dependent NAD(+) synthetase Pseudomonas aeruginosa in complex with NAD

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	100
Detector technology	CCD
Collection date	2015-10-20
Detector	RIGAKU SATURN 944+
Wavelength(s)	1.5418
Spacegroup name	C 1 2 1
Unit cell lengths	111.870, 41.190, 64.130
Unit cell angles	90.00, 113.54, 90.00

Refinement procedure

Resolution	49.712 - 1.500
R-factor	0.1404
Rwork	0.138
R-free	0.17990
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4xfd apo structure
RMSD bond length	0.007
RMSD bond angle	0.899
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHENIX ((dev_2236))
Refinement software	PHENIX ((dev_2236))

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000		1.540
High resolution limit [Å]	1.500	6.710	1.500
Rmerge	0.039	0.032	0.349
Rmeas	0.040	0.033	0.386
Total number of observations	432740
Number of reflections	42621	527	2952
<I/σ(I)>	31.81	104.3	4.44
Completeness [%]	98.5	99.1	92.2
Redundancy	10.1		5.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	290	Morpheus screen, e1: 10% w/v PEG 20 000, 20% v/v PEG MME 550; 30mM of each diethyleneglycol, triethyleneglycol, tetraethyleneglycol, pentaethyleneglycol; 0.1 M MES/imidazole pH 6.5; PsaeA.00025.a.B1.PW37555 at 23.5 mg/ml; 5 h soak with 5mM NAD; cryo: direct; tray 258195e1, puck hml20-1