5C5I

Crystal structure of NADP-dependent dehydrogenase from Rhodobacter sphaeroides

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	2014-11-06
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97856
Spacegroup name	C 1 2 1
Unit cell lengths	110.048, 80.085, 61.330
Unit cell angles	90.00, 93.69, 90.00

Refinement procedure

Resolution	50.000 - 2.200
R-factor	0.1973
Rwork	0.195
R-free	0.25200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4zqb
RMSD bond length	0.014
RMSD bond angle	1.657
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	2.240
High resolution limit [Å]	2.200	5.970	2.200
Rmerge	0.074	0.042	0.527
Rmeas	0.085	0.048	0.602
Rpim	0.040	0.023	0.286
Total number of observations	113742
Number of reflections	26059
<I/σ(I)>	8.7		2.2
Completeness [%]	96.4	98.2	79.5
Redundancy	4.4	4.1	3.9
CC(1/2)		0.996	0.893

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 # 72 (20%w/v PEG 3350, 0.2M Li nitrate) 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