4WJI

Crystal structure of cyclohexadienyl dehydrogenase from Sinorhizobium meliloti in complex with NADP and tyrosine

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	2013-06-20
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.97856
Spacegroup name	C 1 2 1
Unit cell lengths	76.409, 68.891, 51.097
Unit cell angles	90.00, 94.34, 90.00

Refinement procedure

Resolution	50.000 - 1.400
R-factor	0.1173
Rwork	0.115
R-free	0.15870
Structure solution method	SAD
RMSD bond length	0.014
RMSD bond angle	1.616
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	HKL-3000
Refinement software	REFMAC (5.8.0073)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.420
High resolution limit [Å]	1.400	3.800	1.400
Rmerge	0.066	0.044	0.369
Rmeas	0.084	0.056	0.501
Rpim	0.052	0.034	0.336
Total number of observations	112812
Number of reflections	49774
<I/σ(I)>	14.3		1.8
Completeness [%]	95.6	84.9	93.4
Redundancy	2.3	2.4	1.7
CC(1/2)		0.993	0.734

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	289	0.2 ul of 11 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azid, 0.5 mM TCEP, 7mM NADP and 20 mM tyrosine were mixed with 0.2 ul of the Index condition #83 (0.2 M Magnesium chloride hexahydrate, 0.1 M BIS-TRIS pH 6.5, 25% w/v Polyethylene glycol 3,350) 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 solution at 289 K for 3 hours