8A67
Branched Lys48- and Lys63-linked tri-ubiquitin (K48-K63-Ub3) in complex with matured synthetic nanobody NbSL3.3Q (3rd generation)
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | ESRF BEAMLINE ID23-2 |
Synchrotron site | ESRF |
Beamline | ID23-2 |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2022-06-09 |
Detector | DECTRIS PILATUS3 2M |
Wavelength(s) | 0.87313 |
Spacegroup name | P 1 |
Unit cell lengths | 57.081, 58.243, 61.662 |
Unit cell angles | 78.88, 67.94, 80.16 |
Refinement procedure
Resolution | 52.590 - 1.860 |
R-factor | 0.1893 |
Rwork | 0.186 |
R-free | 0.24520 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 7nbb |
RMSD bond length | 0.002 |
RMSD bond angle | 0.503 |
Data reduction software | autoPROC |
Data scaling software | XSCALE |
Phasing software | PHENIX |
Refinement software | PHENIX (1.20.1_4487) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 56.580 | 2.090 |
High resolution limit [Å] | 1.860 | 1.860 |
Number of reflections | 36500 | 1826 |
<I/σ(I)> | 4.1 | |
Completeness [%] | 88.7 | |
Redundancy | 3.2 | |
CC(1/2) | 0.987 | 0.735 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 7.5 | 293.15 | Protein concentrated to 14.5 mg/ml in 20 mM HEPES pH 7.5, 150 mM NaCl. Mixed 200 nl protein with 100 nl mother liquor (0.1 M HEPES pH 7.5, 10% 2-propanol, 20% PEG4000). Crystals harvested and cryo-protected with Mother liquor supplemented with 30% glycerol. |