5E4B
Hydroxynitrile lyase from the fern Davallia tyermanii in complex with (R)-mandelonitrile / benzaldehyde
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | ESRF BEAMLINE BM14 |
Synchrotron site | ESRF |
Beamline | BM14 |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2014-06-22 |
Detector | MARMOSAIC 225 mm CCD |
Wavelength(s) | 0.9184 |
Spacegroup name | I 2 2 2 |
Unit cell lengths | 73.362, 94.135, 116.143 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 35.800 - 1.500 |
R-factor | 0.1573 |
Rwork | 0.156 |
R-free | 0.17900 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5e46 |
RMSD bond length | 0.007 |
RMSD bond angle | 1.033 |
Data reduction software | XDS |
Data scaling software | XDS |
Phasing software | PHASER |
Refinement software | PHENIX (1.9_1692) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 35.800 | 1.550 |
High resolution limit [Å] | 1.500 | 1.500 |
Rmerge | 0.066 | 0.611 |
Number of reflections | 63070 | |
<I/σ(I)> | 13.74 | 1.86 |
Completeness [%] | 97.5 | 80.88 |
Redundancy | 4.1 | 3 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8 | 298 | Native crystals of DtHNL1 were obtained by mixing 0.5ul 4 mg/mL protein sample (in 10 mM Tris-HCl pH 8.0) with 1 ul reservoir solution (0.9 M NaNO3; Na2HPO4; (NH4)2SO4 mix, 0.1 M Tris-Bicine Buffer pH 8.5 and 30% (w/v) polyethylene glycol monomethyl ether 550 & polyethylene glycol 20k; Morpheus condition C9). Additionally, native crystals were also grown by mixing 1 ul 4 mg/mL protein sample (in 10 mM Tris-HCl pH 8.0) with 0.5ul reservoir solution (0.1 M 2-(4-(2-hydroxyethyl)-1-piperazinyl) ethanesulfonic acid pH 7.5 and 10% (w/v) polyethylene glycol; JSCG condition B4). |