1BKW

p-Hydroxybenzoate hydroxylase (phbh) mutant with cys116 replaced by ser (c116s) and arg44 replaced by lys (r44k), in complex with fad and 4-hydroxybenzoic acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	SIEMENS
Temperature [K]	298
Detector technology	AREA DETECTOR
Collection date	1994-11-15
Detector	SIEMENS
Spacegroup name	C 2 2 21
Unit cell lengths	72.700, 146.400, 88.500
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	8.000 - 2.200
R-factor	0.182
Rwork	0.182
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1pbe
RMSD bond length	0.009
RMSD bond angle	23.500 *
Data reduction software	XDS
Data scaling software	XDS
Phasing software	X-PLOR
Refinement software	X-PLOR (3.8)

Data quality characteristics

	Overall
Low resolution limit [Å]	8.000
High resolution limit [Å]	2.200
Rmerge	0.068 *
Total number of observations	92992 *
Number of reflections	23336
<I/σ(I)>	12
Completeness [%]	96.6
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, 60 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	protein	10 (mg/ml)
2	1	drop	potassium phosphate	100 (mM)	pH7.0
3	1	reservoir	ammonium sulfate	39 (%sat)
4	1	reservoir	FAD	0.14 (mM)
5	1	reservoir	EDTA	0.15 (mM)
6	1	reservoir	4-hydroxybenzoate	1 (mM)
7	1	reservoir	sodium sulfite	60 (mM)
8	1	reservoir	potassium phosphate	100 (mM)	pH7.0