5EAT

5-EPI-ARISTOLOCHENE SYNTHASE FROM NICOTIANA TABACUM WITH SUBSTRATE ANALOG FARNESYL HYDROXYPHOSPHONATE

Experimental procedure

Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL7-1
Synchrotron site	SSRL
Beamline	BL7-1
Temperature [K]	90
Detector technology	IMAGE PLATE
Collection date	1996-11
Detector	MARRESEARCH
Spacegroup name	P 41 21 2
Unit cell lengths	126.330, 126.330, 121.698
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	100.000 - 2.800
R-factor	0.24
Rwork	0.240
R-free	0.28000
Structure solution method	RIGID BODY REFINEMENT/DIFFERENCE FOURIER MAPS
Starting model (for MR)	NATIVE TEAS (PDB ENTRY 5EAS)
RMSD bond length	0.011
RMSD bond angle	19.800 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	X-PLOR (3.1)
Refinement software	X-PLOR (3.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	100.000	2.900
High resolution limit [Å]	2.800	2.800
Rmerge	0.187 *	0.531 *
Total number of observations	136072 *
Number of reflections	24648
<I/σ(I)>	5.4	1.8
Completeness [%]	98.3	96.3
Redundancy	5.5	5.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion, hanging drop *	7 *		PROTEIN WAS CRYSTALLIZED FROM 15% PEG 8000, 200 MM MG(OAC)2, 100 MM MOPSO PH 6.9, 1MM DTT, 2 MM FARNESYL HYDROXYPHOSPHONATE; SOAKING SOLUTION FOR FREEZING ALSO INCLUDED 20% ETHYLENE GLYCOL.

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	reservoir	PEG8000	15 (%)
2	1	reservoir		200 (mM)
3	1	reservoir	MOPSO	100 (mM)
4	1	reservoir	dithiothreitol	1 (mM)