8SH1

Structure of human POT1 DNA binding domain bound to a 5'-phosphorylated junction of a telomeric double-stranded DNA duplex with a 3'-overhang

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]	100
Detector technology	PIXEL
Collection date	2022-06-30
Detector	DECTRIS EIGER2 X 9M
Wavelength(s)	1.12713
Spacegroup name	I 2 2 2
Unit cell lengths	58.470, 170.620, 173.960
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	42.660 - 2.600
R-factor	0.2315
Rwork	0.230
R-free	0.25440
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.003
RMSD bond angle	0.590
Data reduction software	MOSFLM
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.19.2_4158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	86.980	2.720
High resolution limit [Å]	2.600	2.600
Rmerge	0.146
Number of reflections	27182	45826
<I/σ(I)>	10.2
Completeness [%]	99.7
Redundancy	13.4
CC(1/2)	0.997	0.270

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	289	hDBD complex with 5prime-P-ds-ss1-12 was crystallized at 16C by the sitting drop method in a drop containing 0.5 microliter 14 mg/ml protein-DNA complex (in 25 mM Tris (pH 8), 100 mM NaCl, and 2 mM DTT) and 0.5 microliter well solution (0.02 M MgCl2, 0.9 M Hepes (pH 7.5), and 20% polyacrylic acid N100 sodium salt, derived from condition G2 of the JCSG+ screen (NeXtal). Crystals were harvested in 25 mM Tris (pH 8), 100 mM NaCl, 0.02 M MgCl2, 0.1 M Hepes (pH 7.5), and 22% polyacrylic acid N100 sodium salt and cryoprotected in harvesting solution supplemented with 35% ethylene glycol