8FIK
APOBEC3A E72A inactive mutant in complex with ATTC-hairpin DNA substrate
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | AUSTRALIAN SYNCHROTRON BEAMLINE MX2 |
Synchrotron site | Australian Synchrotron |
Beamline | MX2 |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2021-09-28 |
Detector | DECTRIS EIGER X 16M |
Wavelength(s) | 0.953739 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 55.001, 57.229, 92.752 |
Unit cell angles | 90.00, 106.48, 90.00 |
Refinement procedure
Resolution | 48.173 - 1.912 |
Rwork | 0.200 |
R-free | 0.23170 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5keg |
RMSD bond length | 0.008 |
RMSD bond angle | 1.737 |
Data reduction software | XDS |
Data scaling software | Aimless |
Phasing software | MOLREP |
Refinement software | REFMAC (5.8.0267) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 48.173 | 1.960 |
High resolution limit [Å] | 1.910 | 1.910 |
Rmerge | 0.090 | 0.913 |
Rpim | 0.057 | 0.589 |
Number of reflections | 42309 | 2706 |
<I/σ(I)> | 7.5 | 1 |
Completeness [%] | 98.4 | 94.2 |
Redundancy | 3.4 | 3.3 |
CC(1/2) | 0.996 | 0.471 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 6.6 | 285 | A3A-E72A (50 mM MES pH 6.0, 100 mM NaCl, 1 mM TCEP, 0.2 mM EDTA) was mixed with oligonucleotides (10 mM Tris/HCl pH 7.9, 1 mM EDTA) at 0.85 mM and 1.7 mM respectively. Dilution was done with protein buffer. The mixture was added to crystallization liquid 1 to 1 and the mixture was pipetted on siliconized glass disks and sealed on top of a reservoir of crystallization liquid for hanging drop crystallization at 12 degrees Celsius. The crystallization liquid has the following composition: 100 mM Bicine at pH 6.6, 200 mM NaCl, 20 mM putrescine, 1 mM TCEP, 1 mM inositol hex phosphate (phytic acid) and 45 % pentaerythritol propoxylate (5/4 PO/OH) |