4WHR

Anhydride reaction intermediate trapped in Protocatechuate 3,4-dioxygenase (pseudomonas putida) at pH 8.5

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2014-07-11
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.97918
Spacegroup name	I 2 2 2
Unit cell lengths	128.437, 140.644, 168.291
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	31.470 - 1.580
R-factor	0.1506
Rwork	0.148
R-free	0.19180
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3t63
RMSD bond length	0.009
RMSD bond angle	1.465
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.610
High resolution limit [Å]	1.580	4.290	1.580
Rmerge	0.142	0.069	0.351
Rmeas	0.162	0.079	0.454
Rpim	0.076	0.037	0.283
Total number of observations	756615
Number of reflections	188155
<I/σ(I)>	12.2
Completeness [%]	91.4	49.6	93.7
Redundancy	4	4.2	2.1
CC(1/2)		0.985	0.794

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5	295	1.2 M Ammonium Sulfate, 2.5 mM 2-mercaptoethanol, 100 mM Tris-HCl pH 8.5; 2:1 ratio of well sol. to 40 mg/ml protein solution in drop