4LTV

Crystal structure of epi-isozizaene synthase from Streptomyces coelicolor A3(2)

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X29A
Synchrotron site	NSLS
Beamline	X29A
Temperature [K]	100
Detector technology	CCD
Collection date	2012-01-31
Detector	ADSC QUANTUM 315
Wavelength(s)	1.075
Spacegroup name	P 1 21 1
Unit cell lengths	52.238, 46.826, 75.817
Unit cell angles	90.00, 95.37, 90.00

Refinement procedure

Resolution	41.068 - 2.403
R-factor	0.197
Rwork	0.191
R-free	0.24950
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	PDBID: 3KB9
RMSD bond length	0.004
RMSD bond angle	0.754
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX ((phenix.refine: 1.5_2))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.490
High resolution limit [Å]	2.400	2.400
Rmerge	0.091	0.335
Number of reflections	13825
<I/σ(I)>	11.912	2.423
Completeness [%]	95.8	94.5
Redundancy	3.1	2.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	298	4 uL drop of protein solution [8-10 mg/mL EIZS, 20 mM Tris-HCl (pH 7.5), 300 mM NaCl, 10 mM MgCl2, 10% glycerol, 2 mM TCEP, 2 mM sodium pyrophosphate, and 2 mM benzyltriethylammonium chloride] was added to 4 uL of precipitant solution [100 mM Bis-Tris (pH 5.5), 25-28%polyethylene glycol 3350, and 0.2M(NH4)2SO4] and equilibrated against a 1 mL well reservoir of precipitant solution. Crystals appeared within 2-3 days. Then the crystals were soaked in a chelation buffer [5 mM 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, 15 mM ethylenediaminetetraacetic acid, 100 mM Bis-Tris (pH 5.5), 30% polyethylene glycol 3350, 200 mM (NH4)2SO4, 15% glycerol] for 7 days. , VAPOR DIFFUSION, SITTING DROP, temperature 298K