2JIQ
A New Catalytic Mechanism of Periplasmic Nitrate Reductase from Desulfovibrio desulfuricans ATCC 27774 from Crystallographic and EPR Data and based on detailed analysis of the sixth ligand
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ESRF BEAMLINE ID14-1 |
| Synchrotron site | ESRF |
| Beamline | ID14-1 |
| Temperature [K] | 100 |
| Detector technology | CCD |
| Collection date | 2004-07-10 |
| Detector | ADSC CCD |
| Spacegroup name | P 31 2 1 |
| Unit cell lengths | 105.212, 105.212, 130.685 |
| Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
| Resolution | 91.290 - 2.440 |
| R-factor | 0.159 |
| Rwork | 0.155 |
| R-free | 0.24800 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 2nap |
| RMSD bond length | 0.014 |
| RMSD bond angle | 1.604 |
| Phasing software | MOLREP |
| Refinement software | REFMAC (5.2.0019) |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 53.000 | 2.560 |
| High resolution limit [Å] | 2.430 | 2.430 |
| Rmerge | 0.270 | 1.140 |
| Number of reflections | 31747 | |
| <I/σ(I)> | 6.3 | 1 |
| Completeness [%] | 99.0 | 93.6 |
| Redundancy | 4.8 | 3.7 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | 6 | COCRYSTALLISATION WITH 200MM OF NO3 (NITRATE) IN 200MM OF DDNAPA (IE PROTEIN) WHICH HAD BEEN REDUCED BY DITHIONITE IN ANAEROBIC CONDITIONS AND THEN AIR REOXIDISED BEFORE SETTTING UP FOR CRYSTALLISATIONS STRAIGHT AFTER THAWING FROM 193K. SAMPLE WAS DIALYSED AND CONCENTRATED BEFORE CRYSTALLISING IN 0.1M MES PH 6.0, 7.5% PEG 8000. CRYSTAL WAS SOAKED WITH 10MM NANO3 CONTAINING CRYOBUFFER. |
