1BF3

P-HYDROXYBENZOATE HYDROXYLASE (PHBH) MUTANT WITH CYS 116 REPLACED BY SER (C116S) AND ARG 42 REPLACED BY LYS (R42K), IN COMPLEX WITH FAD AND 4-HYDROXYBENZOIC ACID

Experimental procedure

Source type	ROTATING ANODE
Source details	MACSCIENCE M18X
Temperature [K]	298
Detector technology	AREA DETECTOR
Collection date	1995-07
Detector	SIEMENS
Spacegroup name	C 2 2 21
Unit cell lengths	71.350, 145.900, 88.800
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	8.000 - 2.200
R-factor	0.175 *
Rwork	0.175
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1pbe
RMSD bond length	0.009
RMSD bond angle	23.800 *
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	X-PLOR (3.8)
Refinement software	X-PLOR (3.8)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	8.000	2.000
High resolution limit [Å]	2.200	1.700
Rmerge	0.067 *	0.045
Total number of observations	78553 *
Number of reflections	23174
<I/σ(I)>		3
Completeness [%]	96.8	82
Redundancy	2.5	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	39 (%sat)
4	1	reservoir	FAD	0.04 (mM)
5	1	reservoir	EDTA	0.15 (mM)
6	1	reservoir	sodium sulfite	30 (mM)
7	1	reservoir	4-hydroxybenzoate	1 (mM)
8	1	reservoir	potassium phosphate	100 (mM)