4GHF

Structure of Y257F variant of Homoprotocatechuate 2,3-Dioxygenase from B.fuscum in complex with 4-Nitrocatechol and dioxygen at 1.67 Ang resolution

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X10SA
Synchrotron site	SLS
Beamline	X10SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2010-05-17
Detector	PSI PILATUS 6M
Wavelength(s)	0.9999
Spacegroup name	P 21 21 2
Unit cell lengths	110.519, 150.318, 96.105
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	89.040 - 1.670
R-factor	0.15236
Rwork	0.151
R-free	0.18052
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3ojt
RMSD bond length	0.015
RMSD bond angle	1.616
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	REFMAC (5.6.0117)
Refinement software	REFMAC (5.6.0117)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	96.110	1.760
High resolution limit [Å]	1.670	1.670
Rmerge	0.035	0.427
Number of reflections	184533
<I/σ(I)>	13.5	2
Completeness [%]	99.6
Redundancy	4.7	4.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	13% PEG6000, 0.1M calcium chloride, 0.1M Tris-HCl. Cryoprotectant 25% PEG400. Ligand soaking: 2mM 4-nitrocatechol under aerobic conditions for 1hr prior to cryo-cooling in liquid nitrogen. , pH pH7.0, VAPOR DIFFUSION, HANGING DROP, temperature 293K