1BGJ

P-HYDROXYBENZOATE HYDROXYLASE (PHBH) MUTANT WITH CYS 116 REPLACED BY SER (C116S) AND HIS 162 REPLACED BY ARG (H162R), IN COMPLEX WITH FAD AND 4-HYDROXYBENZOIC ACID

Experimental procedure

Source type	ROTATING ANODE
Source details	MACSCIENCE M18X
Temperature [K]	280
Detector technology	AREA DETECTOR
Collection date	1995-12
Detector	SIEMENS
Spacegroup name	C 2 2 21
Unit cell lengths	71.800, 146.000, 88.300
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	8.000 - 3.000
R-factor	0.128
Rwork	0.128
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1pbe
RMSD bond length	0.010
RMSD bond angle	24.300 *
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	X-PLOR (3.8)
Refinement software	X-PLOR (3.8)

Data quality characteristics

	Overall
Low resolution limit [Å]	8.000
High resolution limit [Å]	3.000
Rmerge	0.130 *
Number of reflections	6695
Completeness [%]	72.4
Redundancy	2.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7	4 *	39% AMMONIUMSULFATE, 100 MM SODIUM PHOSPHATE, 0.04 MM FAD, 0.15 MM EDTA, 30 MM SODIUM SULFITE, 1 MM P-HYDROXYBENZOATE, pH 7.0

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	enzyme	10-15 (mg/ml)
2	1	drop	potassium phosphate	100 (mM)
3	1	reservoir	ammonium sulfate	40 (%sat)
4	1	reservoir	FAD	0.04 (mM)
5	1	reservoir	EDTA	0.15 (mM)
6	1	reservoir	POHB	2 (mM)
7	1	reservoir	sodium sulfite	30 (mM)
8	1	reservoir	potassium phosphate	100 (mM)