5UDR

LarE, a sulfur transferase involved in synthesis of the cofactor for lactate racemase in complex with nicotinamide mononucleotid NMN

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2016-11-25
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.9786
Spacegroup name	P 41 2 2
Unit cell lengths	108.095, 108.095, 324.964
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.420 - 2.620
R-factor	0.217
Rwork	0.215
R-free	0.24650
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	0.733
Data reduction software	XDS
Data scaling software	Aimless (0.5.29)
Phasing software	PHASER (2.7.16)
Refinement software	PHENIX (1.11.1-2575)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	48.420	48.420	2.690
High resolution limit [Å]	2.620	11.410	2.620
Rmerge	0.098	0.032	1.286
Number of reflections	59143
<I/σ(I)>	21.3
Completeness [%]	99.9	98.6	99
Redundancy	12.2	8.9	12.1
CC(1/2)	0.999	0.999	0.720

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	294.15	5 ul 23 mg/ml LarE (100 mM Tris-HCl pH 7.5, 300 mM NaCl) was mixed with 5 ul of reservoir solution. Hanging drop reservoir contained 100 ul of 30.0% v/v pentaerythritol ethoxylate (15/4 EO/OH), 50 mM BIS-TRIS pH 6.5, 150 mM ammonium sulfate. Crystals grew to full size within 4 days and remained in the drop for 4 months without notable changes. A single crystal was then transfered to a 10 ul drop containing ~100 mM beta-nicotinamide mononucleotide, 30.0% v/v pentaerythritol ethoxylate (15/4 EO/OH) and 50 mM BIS-TRIS pH 6.5. The drop was incubated for 11 days over a hanging drop reservoir containing 100 ul of 30.0% v/v pentaerythritol ethoxylate (15/4 EO/OH), 50 mM BIS-TRIS pH 6.5, 100 mM ammonium sulfate before being directly frozen.