5BWE

Benzylsuccinate synthase alpha-beta-gamma complex with bound toluene and fumarate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X26C
Synchrotron site	NSLS
Beamline	X26C
Temperature [K]	100
Detector technology	CCD
Collection date	2011-07-19
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.00000
Spacegroup name	P 21 21 2
Unit cell lengths	141.534, 115.360, 121.701
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	38.270 - 3.300
R-factor	0.2133
Rwork	0.212
R-free	0.23660
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.003
RMSD bond angle	0.913
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	PHENIX (1.9_1692)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000	3.360
High resolution limit [Å]	3.300	3.300
Rmerge		1.000
Number of reflections	30311
<I/σ(I)>	9.6	1.3
Completeness [%]	98.9	95.4
Redundancy	3.8	3.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	291	2:1 8 mg/mL BSS-alpha-beta-gamma (in 50 mM Tris, pH 7.6, 15% v/v glycerol, 200 mM sodium chloride) to well solution (25% w/v PEG3350, 100 mM Tris, pH 8.5, 60 mM potassium chloride, 5 mM fumarate), 1-2 uL toluene was added to the bottom of the well and allowed to diffuse slowly into the protein drop, diffraction-quality crystals grew over the course of three weeks in a temperature-controlled anaerobic chamber