4WCZ

Crystal structure of a putative enoyl-CoA hydratase/isomerase from Novosphingobium aromaticivorans

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-BM
Synchrotron site	APS
Beamline	19-BM
Temperature [K]	100
Detector technology	CCD
Collection date	2013-11-14
Detector	ADSC QUANTUM 210r
Wavelength(s)	0.97936
Spacegroup name	P 1 21 1
Unit cell lengths	81.636, 128.469, 84.467
Unit cell angles	90.00, 116.36, 90.00

Refinement procedure

Resolution	50.000 - 1.820
R-factor	0.1685
Rwork	0.167
R-free	0.19260
Structure solution method	SAD
RMSD bond length	0.005
RMSD bond angle	1.008
Data reduction software	DENZO
Data scaling software	HKL-3000
Phasing software	SHELXD
Refinement software	REFMAC (5.8.0073)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.850
High resolution limit [Å]	1.820	4.940	1.820
Rmerge	0.104	0.060
Rmeas	0.114	0.068
Rpim	0.054	0.031	0.598
Total number of observations	654357
Number of reflections	138544
<I/σ(I)>	10
Completeness [%]	99.5	99.4	93.8
Redundancy	4.7	4.6	3.6
CC(1/2)		0.995	0.490

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	289	0.2 ul of 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 Hampton Index HT #65 (F5) (0.1 M Ammonium acetate, 0.1 M BIS_TRIS pH 5.5, 17% (w/v) Polyethylene glycol 10,000) and equilibrated against 1.25 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.