4ZQB

Crystal structure of NADP-dependent dehydrogenase from Rhodobactersphaeroides in complex with NADP and sulfate

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	2014-07-23
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97856
Spacegroup name	P 61
Unit cell lengths	71.489, 71.489, 244.364
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	50.000 - 1.850
R-factor	0.1665
Rwork	0.165
R-free	0.19970
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3pp8
RMSD bond length	0.012
RMSD bond angle	1.565
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	HKL-3000
Refinement software	REFMAC (5.8.0107)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.880
High resolution limit [Å]	1.850	5.020	1.850
Rmerge	0.161	0.065
Rmeas	0.171	0.072
Rpim	0.073	0.030	0.428
Total number of observations	368480
Number of reflections	59896
<I/σ(I)>	4.7		2
Completeness [%]	99.3	97.3	97.1
Redundancy	6.2	5.5	6.2
CC(1/2)		0.995	0.649

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	289	0.2 ul of 10.5 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide and 0.5 mM TCEP were mixed with 0.2 ul of the MCSG Suite 2 condition # 23 (0.1M Bis-Tris, 25%w/v PEG 3350, 0.2M Li sulfate pH=6.5 ) and equilibrated against 1.5 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci). Before crystallization protein was incubated with 1/50 v/v of 2 mg/ml chymotrypsin and 10 mM NADP solution at 289 K for 3 hours.