8DF7

Structure of M. kandleri topoisomerase V in complex with DNA. 38 base pair symmetric 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	2019-08-13
Detector	RAYONIX MX-300
Wavelength(s)	0.97872
Spacegroup name	P 43 21 2
Unit cell lengths	121.665, 121.665, 498.820
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	59.100 - 3.520
R-factor	0.2313
Rwork	0.229
R-free	0.27480
Structure solution method	SIRAS
RMSD bond length	0.002
RMSD bond angle	0.427
Data reduction software	XDS
Data scaling software	STARANISO
Phasing software	SHARP
Refinement software	BUSTER

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	59.100	3.730
High resolution limit [Å]	3.520	3.520
Rmerge	0.121	1.183
Rmeas	0.128	1.289
Number of reflections	41745	13495
<I/σ(I)>	12	1.7
Completeness [%]	87.5	28.2
Redundancy	9.5	6.5
CC(1/2)	0.997	0.633

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 5.5, 12.5 uM phosphotungstic acid