1W54

Stepwise introduction of a zinc binding site into Porphobilinogen synthase from Pseudomonas aeruginosa (mutation D139C)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RUH3R
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2002-03-15
Detector	RIGAKU R-AXIS IV
Spacegroup name	P 4 21 2
Unit cell lengths	125.231, 125.232, 86.030
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	87.710 - 2.200
R-factor	0.157
Rwork	0.154
R-free	0.21700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1b4k
RMSD bond length	0.026
RMSD bond angle	1.928
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	REFMAC (5.1.24)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	47.100	2.280
High resolution limit [Å]	2.200	2.200
Rmerge	0.100	0.450
Number of reflections	35228
<I/σ(I)>	23.1	5.3
Completeness [%]	99.8	99.5
Redundancy	12.6	11.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5		HANGING DROP IN 24-WELL LIMBRO PLATES. DROPS MADE OF 5 MICROL PROTEIN SOLUTION (8,7 MG/ML PROTEIN, 50 MM NA-HEPES PH 7.5, 10 MM MGCL2, 10 ZNCL2, 10 MM DTT) PLUS 5 MICROL RESERVOIR SOLUTION (28,5 % (W/V) PEG-400, 100 MM NA-HEPES PH 7.5, 80MM MGCL2, 20MM DTT) ABOVE 500 MICROL OF RESERVOIR SOLUTION.