3T63

Axial Ligand Swapping In Double Mutant Maintains Intradiol-cleavage Chemistry in Protocatechuate 3,4-Dioxygenase

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	2009-10-24
Detector	ADSC QUANTUM 210r
Wavelength(s)	0.98
Spacegroup name	I 2 2 2
Unit cell lengths	128.119, 140.618, 167.879
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.090 - 1.540
R-factor	0.1562
Rwork	0.155
R-free	0.17780
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2PCD CHAINS A/M B/N C/O
RMSD bond length	0.011
RMSD bond angle	1.346
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	MOLREP
Refinement software	REFMAC

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.570
High resolution limit [Å]	1.540	4.180	1.540
Rmerge	0.049	0.028	0.423
Number of reflections	218848
<I/σ(I)>	18.6
Completeness [%]	99.1	94.7	98.5
Redundancy	5.5	5.2	5.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5	277	1.5-1.8 M ammonium sulfate, 5 mM BME, 40-60 mM Tris, pH 8.5, varying ML:ENZ ratios from 1:2 to 4:1, VAPOR DIFFUSION, HANGING DROP, temperature 277K