5X8F

Ternary complex structure of a double mutant I454RA456K of o-Succinylbenzoate CoA Synthetase (MenE) from Bacillus Subtilis bound with AMP and its product analogue OSB-NCoA at 1.76 angstrom

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRF BEAMLINE BL17U
Synchrotron site	SSRF
Beamline	BL17U
Temperature [K]	100
Detector technology	CCD
Collection date	2016-09-11
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.979
Spacegroup name	P 1
Unit cell lengths	71.870, 96.360, 98.070
Unit cell angles	80.44, 77.96, 81.11

Refinement procedure

Resolution	29.957 - 1.763
R-factor	0.1635
Rwork	0.163
R-free	0.19350
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5buq
RMSD bond length	0.006
RMSD bond angle	0.883
Data reduction software	Aimless (0.5.26)
Data scaling software	Aimless (0.5.26)
Phasing software	PHENIX (1.10.1_2155)
Refinement software	PHENIX (1.10.1_2155)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	36.990	36.990	1.790
High resolution limit [Å]	1.760	9.660	1.760
Rmerge	0.047	0.029	0.372
Rmeas	0.066	0.041	0.525
Rpim	0.047	0.029	0.372
Number of reflections	230857		11218
<I/σ(I)>	5.8		1.9
Completeness [%]	92.1	52.6	90.8
Redundancy	2	1.8	3.9
CC(1/2)	0.996	0.992	0.827

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.7	289	Reservoir solution: 32% PEG300, 0.1M cacodylate pH6.7, 0.2M Ca(OAc)2, protein solution: 2.5mM OSBNCoA, 0.8mM AMP, 19mg/mL bsMenE I454RA456K, incubated for 4days