4LUU

V329A Epi-isozizaene synthase: Complex with Mg, inorganic pyrophosphate and benzyl triethyl ammonium cation

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	2010-03-04
Detector	ADSC QUANTUM 315
Wavelength(s)	1.008
Spacegroup name	P 1 21 1
Unit cell lengths	53.214, 47.485, 75.283
Unit cell angles	90.00, 95.50, 90.00

Refinement procedure

Resolution	32.797 - 1.950
R-factor	0.1557
Rwork	0.152
R-free	0.20050
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	PDBID: 3KB9
RMSD bond length	0.007
RMSD bond angle	1.014
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASES
Refinement software	PHENIX ((phenix.refine: 1.6_289))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.020
High resolution limit [Å]	1.950	1.950
Rmerge	0.116	0.432
Number of reflections	26839
<I/σ(I)>	12.957	3.029
Completeness [%]	97.3	97.4
Redundancy	4.2	3.7

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 V329A 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