4XCV
Probable 2-hydroxyacid dehydrogenase from Rhizobium etli CFN 42 in complex with NADPH
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | APS BEAMLINE 19-ID |
Synchrotron site | APS |
Beamline | 19-ID |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2014-07-23 |
Detector | ADSC QUANTUM 315r |
Wavelength(s) | 0.97912 |
Spacegroup name | P 41 21 2 |
Unit cell lengths | 65.663, 65.663, 151.458 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 50.000 - 1.400 |
R-factor | 0.1507 |
Rwork | 0.149 |
R-free | 0.17750 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 3kbo |
RMSD bond length | 0.014 |
RMSD bond angle | 1.748 |
Data scaling software | HKL-3000 |
Phasing software | HKL-3000 |
Refinement software | REFMAC (5.8.0073) |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 50.000 | 50.000 | 1.420 |
High resolution limit [Å] | 1.400 | 3.800 | 1.400 |
Rmerge | 0.067 | 0.038 | 0.914 |
Rmeas | 0.072 | 0.040 | 0.986 |
Rpim | 0.025 | 0.014 | 0.361 |
Total number of observations | 426874 | ||
Number of reflections | 62557 | ||
<I/σ(I)> | 10.3 | 2.1 | |
Completeness [%] | 94.8 | 91.4 | 89 |
Redundancy | 6.8 | 8.6 | 6.4 |
CC(1/2) | 0.998 | 0.777 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8.5 | 289 | 0.2 ul of 15 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide, 0.5 mM TCEP 10 mM NADP and 10 mM glycolic acid were mixed with 0.2 ul of the MCSG-I condition #45 (0.2 M Sodium Chloride, 0.1 M Tris:HCl pH 8.5, 25% (w/v) PEG 3350) and equilibrated against 1.5 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci). Before crystallization the protein was incubated with 1/50 v/v of 2 mg/ml TEV solution at 289 K. |