4LXW

L72V 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	52.977, 47.236, 75.084
Unit cell angles	90.00, 95.66, 90.00

Refinement procedure

Resolution	41.203 - 2.092
R-factor	0.1758
Rwork	0.173
R-free	0.20580
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	PDBID: 3KB9
RMSD bond length	0.002
RMSD bond angle	0.649
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	2.180
High resolution limit [Å]	2.092	2.100
Rmerge	0.104	0.301
Number of reflections	22011
<I/σ(I)>	12.204	3.479
Completeness [%]	99.9	99.2
Redundancy	3.6	3.5

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 L72V 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 room temperature., VAPOR DIFFUSION, HANGING DROP, temperature 298K