1AMU

PHENYLALANINE ACTIVATING DOMAIN OF GRAMICIDIN SYNTHETASE 1 IN A COMPLEX WITH AMP AND PHENYLALANINE

Experimental procedure

Source type	SYNCHROTRON
Source details	EMBL/DESY, HAMBURG BEAMLINE BW7B
Synchrotron site	EMBL/DESY, HAMBURG
Beamline	BW7B
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1996-09-09
Detector	MAR scanner 300 mm plate
Spacegroup name	P 1 21 1
Unit cell lengths	61.680, 154.770, 65.300
Unit cell angles	90.00, 93.91, 90.00

Refinement procedure

Resolution	20.000 - 1.900
R-factor	0.213
Rwork	0.213
R-free	0.24600
Structure solution method	MIR
RMSD bond length	0.008
RMSD bond angle	24.100 *
Data reduction software	MOSFLM
Data scaling software	CCP4
Phasing software	X-PLOR (3.1)
Refinement software	X-PLOR (3.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.000
High resolution limit [Å]	1.900	1.900
Rmerge	0.051	0.232
Total number of observations	230285 *
Number of reflections	91144
<I/σ(I)>	7.3	2.7
Completeness [%]	96.4	88.3
Redundancy	2.5	1.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7.8 *	18 *	CRYSTALS WERE GROWN USING THE VAPOR DIFFUSION TECHNIQUE. THE PROTEIN AT 15 MG/ML IN 10 MM TRIS-HCL PH7.8, 50 MM NACL, 15% GLYCEROL, 2 MM ATP, 2 MM L-PHE AND 4 MM MGCL2 WAS EQUILIBRATED WITH AN EQUAL VOLUME OF RESERVOIR SOLUTION CONTAINING 28-32%(W/V) METHOXY POLYETHYLENE GLYCOL 5000, 200 MM AMMONIUM SULFATE, AND 100 MM ADA PH6.5 AT 18C., vapor diffusion, temperature 291K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	15 (mg/ml)
10	1	reservoir	ADA	100 (mM)
2	1	drop	Tris-HCl	10 (mM)
3	1	drop		50 (mM)
4	1	drop	glycerol	15 (%)
5	1	drop	ATP	2 (mM)
6	1	drop	L-phenylalanine	2 (mM)
7	1	drop		4 (mM)
8	1	reservoir	MePEG5000	28-32 (%(w/v))
9	1	reservoir	ammonium sulfate	200 (mM)