8UN9

Crystal structure of the MrfB exonuclease catalytic core

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-D
Synchrotron site	APS
Beamline	21-ID-D
Temperature [K]	289
Detector technology	PIXEL
Collection date	2023-02-07
Detector	DECTRIS EIGER X 9M
Wavelength(s)	1.127129
Spacegroup name	P 1 21 1
Unit cell lengths	72.874, 37.928, 92.616
Unit cell angles	90.00, 99.15, 90.00

Refinement procedure

Resolution	41.740 - 2.103
Rwork	0.175
R-free	0.23250
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.006
RMSD bond angle	1.301
Data reduction software	DIALS
Data scaling software	DIALS
Phasing software	PHASER
Refinement software	REFMAC (5.8.0419)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	45.720	2.160
High resolution limit [Å]	2.100	2.100
Rmerge	0.064	0.489
Rmeas	0.076	0.576
Rpim	0.040	0.303
Number of reflections	29515	2294
<I/σ(I)>	14.2	2.7
Completeness [%]	99.6	97.7
Redundancy	6.5	6.7
CC(1/2)	0.998	0.917

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	289	The MrfB catalytic core (33-279) was crystallized by hanging drop with 1 microliter of 10 mg/mL protein (in 25 mM Tris pH 7.5, 125 mM NaCl, 1 mM TCEP, 5 mM MgCl2, 2 mM dTMP) and 1 microliter of well solution (0.1 M HEPES pH 7.5, 15% PEG Smear medium, 8% ethylene glycol). Crystals were harvested in 20 mM Tris pH 7.5, 125 mM NaCl, 1 mM TCEP, 5 mM MgCl2, 2 mM dTMP, 0.1 M HEPES pH 7.5, 20% PEG smear medium, and cryoprotected with the addition of 25% ethylene glycol