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 |