8DFB

Structure of M. kandleri topoisomerase V in complex with DNA. 39 base pair symmetric DNA complex

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	2021-02-05
Detector	RAYONIX MX-300
Wavelength(s)	0.97872
Spacegroup name	P 43 21 2
Unit cell lengths	121.560, 121.560, 497.191
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	59.040 - 3.170
R-factor	0.2202
Rwork	0.218
R-free	0.25130
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	D_1000262487
RMSD bond length	0.002
RMSD bond angle	0.441
Data reduction software	XDS
Data scaling software	STARANISO
Phasing software	PHASER
Refinement software	PHENIX ((1.19.1_4122: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	86.900	3.360
High resolution limit [Å]	3.170	3.170
Rmerge	0.079	1.330
Rmeas	0.084	1.410
Number of reflections	53311	2667
<I/σ(I)>	18.1	1.5
Completeness [%]	82.4	25.7
Redundancy	8.8	9.9
CC(1/2)	1.000	0.646

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.1	303	Protein was mixed with the annealed oligonucleotide using a stoichiometric ratio of 1.25:1 DNA to protein in 1X DNA binding buffer. Reactions were incubated for thirty minutes at 65 C. Crystals started to appear within minutes of setting up the trays in 1:1 or 2:1 well to complex ratio. Well solution: 2% PEG 8K, 24 mM sodium acetate pH 5.1, 26 mM sodium acetate pH 5.6, 12.5 uM phosphotungstic acid