6TP8
Substrate protein interactions in the limbus region of the catalytic site of Candida antarctica Lipase B
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | EMBL/DESY, HAMBURG BEAMLINE X13 |
Synchrotron site | EMBL/DESY, HAMBURG |
Beamline | X13 |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2013-09-22 |
Detector | DECTRIS PILATUS 6M |
Wavelength(s) | 0.824 |
Spacegroup name | C 2 2 21 |
Unit cell lengths | 89.426, 156.654, 138.084 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 67.690 - 1.550 |
R-factor | 0.1644 |
Rwork | 0.163 |
R-free | 0.19210 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 1tca |
RMSD bond length | 0.006 |
RMSD bond angle | 0.904 |
Data reduction software | XDS |
Data scaling software | Aimless |
Phasing software | MOLREP |
Refinement software | PHENIX (1.13_2998) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 67.690 | 1.605 |
High resolution limit [Å] | 1.550 | 1.550 |
Rmerge | 0.121 | 1.460 |
Rmeas | 0.140 | 1.700 |
Rpim | 0.050 | 0.877 |
Number of reflections | 138806 | 48047 |
<I/σ(I)> | 8.4 | 1 |
Completeness [%] | 99.2 | 98.05 |
Redundancy | 7.6 | 7.2 |
CC(1/2) | 0.997 | 0.550 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 4.8 | 293 | Crystallization trials were performed at 293 K using the hanging-drop method using a Qiagen EasyXtal 15-well plate. A 15 mg.mL CALB solution in 20mM Na(CH3COO) pH = 4.8 was incubated with a solution of paraoxon-ethyl (SIGMA catalog n. N12816) 30mM in water for one day. 1 uL of CALB solution was diluted with 1 uL of the precipitant solution, made of 200mM Na(CH3COO) pH = 4.8, 20% (w.v) PEG4000, and 15% (v.v) glyceryl tributyrate (SIGMA catalog n. T8626). The drop was equilibrated by vapor diffusion against 500 uL of the precipitant solution. Protein crystals of the CALB complex appeared within two weeks and grew to a size of 0.1 x 0.1 x 0.05 mm3 in five weeks. |