5VOH
Crystal structure of engineered water-forming NADPH oxidase (TPNOX) bound to NADPH. The G159A, D177A, A178R, M179S, P184R mutant of LbNOX.
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ALS BEAMLINE 8.2.2 |
| Synchrotron site | ALS |
| Beamline | 8.2.2 |
| Temperature [K] | 100 |
| Detector technology | CCD |
| Collection date | 2016-06-04 |
| Detector | ADSC QUANTUM 315r |
| Wavelength(s) | 0.9999 |
| Spacegroup name | P 1 21 1 |
| Unit cell lengths | 107.264, 96.527, 119.061 |
| Unit cell angles | 90.00, 116.76, 90.00 |
Refinement procedure
| Resolution | 47.888 - 2.302 |
| R-factor | 0.1942 |
| Rwork | 0.192 |
| R-free | 0.25510 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 5er0 |
| RMSD bond length | 0.002 |
| RMSD bond angle | 0.525 |
| Data reduction software | HKL-2000 |
| Data scaling software | HKL-2000 |
| Phasing software | PHASER |
| Refinement software | PHENIX (1.10.1_2155) |
Data quality characteristics
| Overall | Inner shell | Outer shell | |
| Low resolution limit [Å] | 50.000 | 50.000 | 2.340 |
| High resolution limit [Å] | 2.300 | 6.240 | 2.300 |
| Rmerge | 0.102 | 0.037 | 0.560 |
| Rmeas | 0.109 | 0.039 | 0.604 |
| Rpim | 0.040 | 0.014 | 0.225 |
| Total number of observations | 718655 | ||
| Number of reflections | 96175 | ||
| <I/σ(I)> | 6.9 | ||
| Completeness [%] | 100.0 | 99.9 | 99.9 |
| Redundancy | 7.5 | 7.6 | 7.1 |
| CC(1/2) | 0.999 | 0.933 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, HANGING DROP | 6.5 | 294 | Crystals were grown in 25 % PEG 3350, 0.1 M Bis-Tris, pH 6.5, 0.2 M (NH4)2SO4 for TPNOX at 21 oC. Crystals were briefly soaked for 1-3 min in the formulation with 15 % (v/v) of ethylene glycol supplemented with 20-30 mM NADPH and were quickly frozen in liquid nitrogen. |
