1R6W

Crystal structure of the K133R mutant of o-Succinylbenzoate synthase (OSBS) from Escherichia coli. Complex with SHCHC

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 14-BM-D
Synchrotron site	APS
Beamline	14-BM-D
Temperature [K]	150
Wavelength(s)	0.9790
Spacegroup name	P 21 21 2
Unit cell lengths	72.200, 82.900, 56.200
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	30.000 - 1.620
R-factor	0.1691
Rwork	0.167
R-free	0.20000 *
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1fhv
RMSD bond length	0.016
RMSD bond angle	1.620 *
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	MOLREP
Refinement software	REFMAC (5.1.24)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	31.300 *	1.680
High resolution limit [Å]	1.620	1.620
Rmerge	0.046	0.293
Total number of observations	307914 *
Number of reflections	43694
<I/σ(I)>	41	17.1
Completeness [%]	99.8 *	99.9 *
Redundancy	7.05 *

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Batch method *	5.5	20 *	The K133R mutant of OSBS was concentrated to 15 mg/ml, dialyzed against 5 mM Tris pH 8.3 containing 2 mM MgCl2, drop frozen as small pellets in liquid nitrogen and stored at -80 C. Crystals were grown at 20 C by small-scale batch experiments by combining 15 ml of protein solution and 15 ml of a solution containing 12-13% MePEG 5000, 100 mM sodium acetate, 60 mM MgCl2, at pH 5.5. SHCHC was included in the crystallization at a final concentration of approximately 2.5 mM., microbatch, temperature 293K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	1	protein	15 (mg/ml)
2	1	1	Tris-HCl	5 (mM)	pH8.3
3	1	1		2 (mM)
4	1	1	MePEG5000	12-13 (%)
5	1	1	sodium acetate	100 (mM)
6	1	1		60 (mM)	pH5.5