6SMY

Crystal structure of SLA Reductase YihU from E. Coli with NADH and product DHPS

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04-1
Synchrotron site	Diamond
Beamline	I04-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-07-14
Detector	DECTRIS PILATUS 2M
Wavelength(s)	0.9159
Spacegroup name	P 21 21 21
Unit cell lengths	70.121, 113.168, 132.393
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	86.020 - 2.450
R-factor	0.2322
Rwork	0.230
R-free	0.27010
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6sm7
RMSD bond length	0.007
RMSD bond angle	1.423
Data reduction software	xia2
Data scaling software	Aimless (0.7.4)
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0253)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	86.020	2.550
High resolution limit [Å]	2.450	2.450
Rmerge	0.067	0.207
Rmeas	0.071	0.221
Rpim	0.025	0.076
Total number of observations	320728
Number of reflections	39554	4387
<I/σ(I)>	18.2
Completeness [%]	100.0	100
Redundancy	8.1	8.2
CC(1/2)	0.999	0.986

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	YihU was co-crystallized with 5mM NADH using 0.2 M Ammonium acetate, 0.1 M Bis-Tris pH 6.5, 32% PEG 3,350 as precipitant. The crystal was then soaked with solid DHPS in the mother liquor for 2 min prior to harvesting into liquid nitrogen without any cryoprotectant.