5KTN

Crystal structure of Pyrococcus horikoshii quinolinate synthase (NadA) with bound dihydroxyacetone phosphate (DHAP) and Fe4S4 cluster

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-E
Synchrotron site	APS
Beamline	24-ID-E
Temperature [K]	100
Detector technology	CCD
Collection date	2013-04-06
Detector	ADSC QUANTUM 315
Wavelength(s)	0.97925
Spacegroup name	P 1 21 1
Unit cell lengths	47.303, 52.522, 55.544
Unit cell angles	90.00, 112.20, 90.00

Refinement procedure

Resolution	33.636 - 1.340
R-factor	0.1243
Rwork	0.123
R-free	0.15460
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Crystal structure of Pyrococcus horikoshii NadA with bound malate and lacking Fe4S4 cluster (PDB entry 1WZU)
RMSD bond length	0.012
RMSD bond angle	1.728
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX ((1.10_2155: 000))
Refinement software	PHENIX ((1.10_2155: 000))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	35.000	1.390
High resolution limit [Å]	1.340	1.340
Rmerge	0.047	0.242
Number of reflections	56267
<I/σ(I)>	19.3	5
Completeness [%]	99.4	95.1
Redundancy	3.6	2.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION		296	210 mM ammonium chloride, 8 - 15% polyethylene glycol 4000, and 40 mM HEPES, pH 5.5 - 7.5. DHAP was added to the protein solution to a final concentration of 7-10 mM