8DF9

Structure of M. kandleri topoisomerase V in complex with DNA. 38 base pair asymmetric DNA complex

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2018-03-14
Detector	RAYONIX MX-300
Wavelength(s)	0.97856
Spacegroup name	P 41 21 2
Unit cell lengths	193.751, 193.751, 245.979
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	39.280 - 3.240
R-factor	0.2362
Rwork	0.235
R-free	0.26560
Structure solution method	SIRAS
RMSD bond length	0.004
RMSD bond angle	0.562
Data reduction software	XDS
Data scaling software	STARANISO
Phasing software	SHARP
Refinement software	BUSTER

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	39.300	3.601
High resolution limit [Å]	3.240	3.240
Rmerge	0.093	1.480
Rmeas	0.100	1.567
Number of reflections	52587	2629
<I/σ(I)>	13.5	1.7
Completeness [%]	70.4	13.2
Redundancy	8.2	9.4
CC(1/2)	0.999	0.580

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5.5	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: 10% PEG 600, 50 mM sodium succinate pH 5.5, 200 mM potassium chloride, 10 mM magnesium chloride, 1 mM spermine