1CC4

PHE161 AND ARG166 VARIANTS OF P-HYDROXYBENZOATE HYDROXYLASE. IMPLICATIONS FOR NADPH RECOGNITION AND STRUCTURAL STABILITY.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	SIEMENS
Temperature [K]	298
Detector technology	AREA DETECTOR
Collection date	1996-11-15
Detector	SIEMENS
Spacegroup name	C 2 2 21
Unit cell lengths	71.700, 146.300, 88.500
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	8.000 - 2.000
R-factor	0.179 *
Rwork	0.179
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1pbe
RMSD bond length	0.009
RMSD bond angle	24.000 *
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	X-PLOR
Refinement software	X-PLOR (3.851)

Data quality characteristics

	Overall
Low resolution limit [Å]	8.000
High resolution limit [Å]	2.000
Rmerge	0.049 *
Number of reflections	27343
<I/σ(I)>	12
Completeness [%]	94.4
Redundancy	2.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7	4 *	Eppink, M.H.M., (1995) Eur. J. Biochem., 231, 157. *

Crystallization Reagents

ID	crystal ID	solution ID	reagent name	concentration	details
1	1	1	(NH4)2SO4
2	1	1	NA3PO4
3	1	1	NA2SO3
4	1	1	EDTA
5	1	1	P-HYDROXYBENZOATE

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	enzyme	10 (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	4-hydroxybenzoate	1 (mM)
7	1	reservoir	sodium sulfite	60 (mM)
8	1	reservoir	potassium phosphate	100 (mM)