4LZC

W325F Epi-isozizaene synthase: Complex with Mg, inorganic pyrophosphate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	100
Detector technology	CCD
Collection date	2010-06-12
Detector	ADSC QUANTUM 315
Wavelength(s)	0.9795
Spacegroup name	P 1 21 1
Unit cell lengths	53.183, 47.070, 75.585
Unit cell angles	90.00, 95.79, 90.00

Refinement procedure

Resolution	45.487 - 2.457
R-factor	0.1869
Rwork	0.182
R-free	0.23120
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	PDBID: 3KB9
RMSD bond length	0.002
RMSD bond angle	0.575
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASES
Refinement software	PHENIX ((phenix.refine: dev_1391))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.550
High resolution limit [Å]	2.457	2.460
Rmerge	0.148	0.427
Number of reflections	13755
<I/σ(I)>	7.551	3.021
Completeness [%]	99.9	99.9
Redundancy	3.6	3.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.5	298	a 4 uL drop of protein solution [8 mg/mL W325F EIZS, 20 mM Tris-HCl (pH 7.5), 300 mM NaCl, 10 mM MgCl2, 10% glycerol, 1 mM TCEP, 2 mM sodium pyrophosphate, 2 mM benzyltriethylammonium chloride (BTAC)] was added to a 4 uL drop of precipitant solution [100 mM Bis-Tris (pH 5.5), 25-28% polyethylene glycol 3350, 0.2 M (NH4)2SO4] and equilibrated against a 1 mL reservoir of precipitant solution at 298K, VAPOR DIFFUSION, HANGING DROP