4WSA
Crystal structure of Influenza B polymerase bound to the vRNA promoter (FluB1 form)
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | ESRF BEAMLINE ID23-1 |
Synchrotron site | ESRF |
Beamline | ID23-1 |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2014-04-04 |
Detector | DECTRIS PILATUS 6M-F |
Wavelength(s) | 0.9730 |
Spacegroup name | P 32 2 1 |
Unit cell lengths | 199.700, 199.700, 252.680 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 48.978 - 3.400 |
R-factor | 0.2304 |
Rwork | 0.229 |
R-free | 0.26530 |
Structure solution method | SIRAS |
RMSD bond length | 0.003 |
RMSD bond angle | 0.657 |
Data reduction software | XDS |
Data scaling software | XSCALE |
Phasing software | PHASER |
Refinement software | PHENIX ((phenix.refine: dev_1760)) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 50.000 | 3.520 |
High resolution limit [Å] | 3.400 | 3.400 |
Rmerge | 0.121 | 1.300 |
Number of reflections | 79266 | |
<I/σ(I)> | 12.6 | 1.5 |
Completeness [%] | 98.5 | 89.9 |
Redundancy | 11.8 | 5.8 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 9 | 281 | FluB polymerase was concentrated to 9 mg per ml (37 microM) in a buffer containing 500 mM NaCl, 50 mM Hepes pH 7.5, 5% glycerol and 2mM Tris(2-carboxyethyl)phosphine (TCEP), and mixed with 40 microM vRNA for crystallization in hanging drops at 4 C. A trigonal crystal form (FluB1) was obtained by mixing polymerase with nucleotides 5-18 of the 3 prime end and 1-14 of the 5 prime end of the vRNA in a condition containing 0.1 M bicine pH 9.0, 10% MPD. |