3INM
Crystal structure of human cytosolic NADP(+)-dependent isocitrate dehydrogenase R132H mutant in complex with NADPH, ALPHA-KETOGLUTARATE and CALCIUM(2+)
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | NSLS BEAMLINE X29A |
Synchrotron site | NSLS |
Beamline | X29A |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2009-06-13 |
Detector | ADSC QUANTUM 315r |
Wavelength(s) | 1.0809 |
Spacegroup name | C 2 2 21 |
Unit cell lengths | 96.120, 274.690, 116.500 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 24.910 - 2.100 |
R-factor | 0.222 |
Rwork | 0.220 |
R-free | 0.26200 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 1t0l |
RMSD bond length | 0.014 |
RMSD bond angle | 1.105 |
Data reduction software | DENZO |
Data scaling software | SCALEPACK |
Phasing software | PHASER (2.1.4) |
Refinement software | REFMAC |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 25.000 | 2.180 |
High resolution limit [Å] | 2.100 | 2.100 |
Rmerge | 0.080 | 0.305 |
Number of reflections | 83121 | |
<I/σ(I)> | 20.3 | 2.4 |
Completeness [%] | 92.4 | 58.8 |
Redundancy | 8.6 | 2.1 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 6.5 | 293 | Protein component: 8 mg/ml IDH, 20mM Tris-HCl pH 7.5, 100mM sodium chloride, 10mM NADPH, 10mM calcium chloride, 75mM alpha-ketoglutaric acid sodium salt. Precipitant: 100mM MES pH 6.5, 20% PEG 6000. Ratio of protein component to precipitant in initial hanging drop: 2:1., VAPOR DIFFUSION, HANGING DROP, temperature 293K |