4X3G
Crystal structure of SIAH1 SINA domain in complex with a USP19 peptide
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | APS BEAMLINE 19-ID |
Synchrotron site | APS |
Beamline | 19-ID |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2014-07-17 |
Detector | ADSC QUANTUM 315 |
Wavelength(s) | 0.97918 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 41.343, 88.092, 59.590 |
Unit cell angles | 90.00, 103.24, 90.00 |
Refinement procedure
Resolution | 48.450 - 2.340 |
R-factor | 0.2233 |
Rwork | 0.222 |
R-free | 0.25120 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 4i7b |
RMSD bond length | 0.008 |
RMSD bond angle | 0.950 |
Data reduction software | HKL-3000 |
Data scaling software | HKL-3000 |
Phasing software | MOLREP |
Refinement software | BUSTER-TNT (BUSTER 2.10.0) |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 50.000 | 50.000 | 2.370 |
High resolution limit [Å] | 2.330 | 6.320 | 2.330 |
Rmerge | 0.089 | 0.072 | 0.440 |
Total number of observations | 36140 | ||
Number of reflections | 15818 | ||
<I/σ(I)> | 6.2 | ||
Completeness [%] | 90.9 | 87.9 | 58.9 |
Redundancy | 2.3 | 2.4 | 1.5 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 9 | 298 | SIAH1 AT 17.6 MG/ML WAS MIXED WITH A TWO-FOLD EXCESS OF USP19 PEPTIDE ON ICE FOR 30 MIN. BEFORE SETTING UP FOR CRYSTALLIZATION. CRYSTALS WERE GROWN AT 298K USING THE SITTING DROP METHOD BY MIXING 0.5 UL PROTEIN:PEPTIDE MIX WITH 0.5 UL WELL SOLUTION CONSISTING OF 20% PEG6000, 0.1 M BICINE PH 9.0. THE CRYSTALS WERE CRYOPROTECTED BY FIRST IMMERSION IN WELL SOLUTION MIXED WITH 15% (FINAL) ETHYLENE GLYCOL, THEN IMMERSION IN N-PARATONE. |