3PYK
Human Carbonic Anhydrase II as Host for Pianostool Complexes Bearing a Sulfonamide Anchor
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SEALED TUBE |
Source details | OTHER |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2011-04-14 |
Detector | MARMOSAIC 225 mm CCD |
Wavelength(s) | 1.0 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 42.099, 41.493, 72.382 |
Unit cell angles | 90.00, 104.33, 90.00 |
Refinement procedure
Resolution | 30.600 - 1.300 |
R-factor | 0.12947 |
Rwork | 0.128 |
R-free | 0.16471 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 2cba |
RMSD bond length | 0.010 |
RMSD bond angle | 1.454 |
Data reduction software | MOSFLM |
Data scaling software | SCALA |
Phasing software | PHASER |
Refinement software | REFMAC (5.5.0109) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 30.600 | 1.334 |
High resolution limit [Å] | 1.300 | 1.300 |
Rmerge | 0.309 | |
Number of reflections | 59007 | |
<I/σ(I)> | 2.8 | |
Completeness [%] | 96.9 | 91.2 |
Redundancy | 2.7 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 8 | 277 | 3 uL protein solution (20 mg/mL lyophilized hCAII in 50 mM Tris-sulfate, 1 mM methyl mercuric acetate) and 5 uL precipitating buffer (2.6 M ammonium sulfate, 50 mM Tris-sulfate were mixed and equilibrated against 500 uL precipitating buffer. Single crystals were transferred into a cross-linking buffer, which was prepared from 10 uL precipitating buffer and 5 uL glutaraldehyde solution (0.8 % glutaraldehyde, 4 M ammonium sulfate, 50 mM Tris-sulfate) and equilibrated for 18 h. Cross-linked crystals were transferred into soaking buffer (9 uL precipitating buffer, 0.5 uL 60 mM {(6-benzene)Ru(bispy 3)Cl}+ in DMSO) and equilibrated for 54 h, pH 8.0, VAPOR DIFFUSION, HANGING DROP, temperature 277K |