5BPT
Atomic-resolution structures of the APC/C subunits Apc4 and the Apc5 N-terminal domain
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | DIAMOND BEAMLINE I24 |
Synchrotron site | Diamond |
Beamline | I24 |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2013-05-01 |
Detector | ADSC QUANTUM 4 |
Wavelength(s) | 0.969 |
Spacegroup name | P 41 |
Unit cell lengths | 90.469, 90.469, 115.593 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 48.710 - 3.200 |
R-factor | 0.2254 |
Rwork | 0.223 |
R-free | 0.26530 |
Structure solution method | MAD |
RMSD bond length | 0.008 |
RMSD bond angle | 1.070 |
Data reduction software | XDS |
Data scaling software | SCALA |
Phasing software | SHARP |
Refinement software | BUSTER-TNT (2.10.1) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 29.300 | 3.470 |
High resolution limit [Å] | 3.200 | 3.200 |
Rmerge | 0.200 | 0.800 |
Number of reflections | 15477 | |
<I/σ(I)> | 13.3 | 1.6 |
Completeness [%] | 99.8 | 98.5 |
Redundancy | 7.5 | 7.7 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8 | 293 | Apc4 was concentrated to 3.5 mg/ml in a buffer of 20 mM Hepes (pH 8.0), 200 mM NaCl and 2 mM DTT. Initial crystal was obtained by vapour diffusion in setting drop in a buffer containing 0.1 M sodium citrate 5.0, 8% (w/v) PEG 8K. By seeding with the initial crystals, large crystals were grown in a buffer containing 0.1 M sodium citrate 5.0, 3% (w/v) PEG 8K, 250 mM magnesium acetate, 10 mM Tris.HCl (pH 8.5), 200 mM NDSB-211, 4% (v/v) ethylene glycol and 2 mM DTT. |