6BWQ

LarC2, the C-terminal domain of a cyclometallase involved in the synthesis of the NPN cofactor of lactate racemase, in complex with MnCTP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	100
Detector technology	PIXEL
Collection date	2017-11-05
Detector	DECTRIS EIGER X 16M
Wavelength(s)	1.033
Spacegroup name	P 21 3
Unit cell lengths	97.215, 97.215, 97.215
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.608 - 1.850
R-factor	0.1496
Rwork	0.144
R-free	0.19220
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6bwo
RMSD bond length	0.011
RMSD bond angle	0.986
Data reduction software	XDS (2.99)
Data scaling software	Aimless (0.5.32)
Phasing software	PHASER (2.8.0)
Refinement software	PHENIX (1.12-2829)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	48.610	48.610	1.890
High resolution limit [Å]	1.850	9.060	1.850
Rmerge	0.116	0.047	1.044
Rmeas	0.123	0.051	1.111
Rpim	0.043	0.019	0.376
Total number of observations	222377
Number of reflections	26499	264	1623
<I/σ(I)>	11.7
Completeness [%]	100.0	99.5	99.4
Redundancy	8.4	7.2	8.5
CC(1/2)	0.997	0.999	0.615

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	294.15	0.34 ul ~8 mg/ml LarC (100 mM Tris-HCl pH 7.5, 300 mM NaCl) were mixed with 0.17 ul of reservoir solution. Sitting drop reservoir contained 100 ul of 0.1 M MES monohydrate pH 6.5, 12% w/v Polyethylene glycol 20,000. Crystal grew within two months. The crystal was soaked in ~15 mM cytidine triphosphate, ~54 mM manganese chloride + reservoir solution for 28 minutes and then for 2 minutes in 20% Polyethylene glycol 400, 80% reservoir solution.