3V8E

Crystal structure of the yeast nicotinamidase Pnc1p bound to the inhibitor nicotinaldehyde

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	2009-10-12
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.97872
Spacegroup name	H 3
Unit cell lengths	298.717, 298.717, 112.652
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	30.000 - 2.710
R-factor	0.18221
Rwork	0.181
R-free	0.20310
RMSD bond length	0.007
RMSD bond angle	1.204
Refinement software	REFMAC (5.5.0102)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.750
High resolution limit [Å]	2.700	2.700
Number of reflections	101147
<I/σ(I)>		5.3
Completeness [%]	100.0	100
Redundancy	5.7	5.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	298	Protein was dialyzed against buffer containing 15 mM Tris pH 7.5, 50 mM NaCl, 4 mM MgCl2, 10 mM Na Citrate and 5 % glycerol and the inhibitor nicotinaldehyde was added at a 4:1 ratio. The protein (5 mg/ml) was mixed with mother liquor (1.6 M NaOAc, 10 % ethylene glycol, 0.1 M HEPES pH 7.4) at a 1:1 (vol) ratio. Crystals were formed by hanging drop vapor diffusion. Crystals were transferred to a cryoprotectant solution (1.5 M NaOAc, 20% ethylene glycol, 0.1M HEPES pH 7.4) and flash-frozen in liquid nitrogen, VAPOR DIFFUSION, HANGING DROP, temperature 298K