8SH0

Structure of human POT1 DNA binding domain bound to a 5'-phosphorylated junction of a telomeric DNA hairpin 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	2022-07-28
Detector	DECTRIS EIGER2 X 9M
Wavelength(s)	1.12723
Spacegroup name	P 32 2 1
Unit cell lengths	67.230, 67.230, 143.373
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	45.190 - 2.160
R-factor	0.1818
Rwork	0.180
R-free	0.22140
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.003
RMSD bond angle	0.646
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]	71.690	2.230
High resolution limit [Å]	2.160	2.160
Rmerge	0.086	1.131
Number of reflections	20897	36315
<I/σ(I)>	19.6
Completeness [%]	100.0
Redundancy	19.3
CC(1/2)	0.999	0.858

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	289	hDBD complex with 5prime-p-hp-ss1-12 was crystallized at 16C by the hanging drop method in a drop containing 0.8 microliter of 13 mg/ml protein-DNA complex (in 25 mM Tris (pH 8), 100 mM NaCl, and 2 mM DTT) and 0.8 microliter well solution (0.1 M NaOAc (pH 5.2) and 10% PEG 20,000). Crystals were harvested in 25 mM Tris (pH 8), 100 mM NaCl, 0.1 M NaOAc (pH 5.2), and 10% PEG 20,000 and cryoprotected in harvesting solution supplemented with 30% ethylene glycol