6PT8
Crystal Structure of CobT from Methanocaldococcus jannaschii in complex with Adenine Alpha-Ribotide and Nicotinic Acid
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 | PIXEL |
Collection date | 2018-08-15 |
Detector | DECTRIS PILATUS3 X 6M |
Wavelength(s) | 0.97934 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 46.390, 138.640, 51.150 |
Unit cell angles | 90.00, 108.32, 90.00 |
Refinement procedure
Resolution | 41.973 - 1.400 |
R-factor | 0.1542 |
Rwork | 0.153 |
R-free | 0.17480 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 3l0z |
Data reduction software | XDS |
Data scaling software | XSCALE |
Phasing software | PHASER |
Refinement software | PHENIX (1.15.2_3472) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 41.973 | 1.450 |
High resolution limit [Å] | 1.400 | 1.400 |
Rmerge | 0.096 | 0.667 |
Number of reflections | 118689 | 11857 |
<I/σ(I)> | 12.18 | 2.92 |
Completeness [%] | 99.0 | 98.98 |
Redundancy | 6.8 | 6.8 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 298 | Crystals formed by mixing 1:1 ratio of 13.3 mg/mL protein solution with well solution containing 100 mM 4-(2-Hydroxyethyl)-1-piperazinepropanesulfonic acid (HEPPS) pH 8.5 and 22% (w/v) PEG 8K. 1 mM adenine and 50 mM Sodium/Potassium Phosphate pH 7.0 were added to protein prior to mixing with well solution. Substrates were captured in lattice prior to freezing by soaking crystals in 20% (w/v) ethylene glycol, 100 mM HEPPS pH 8.5, 22% (w/v) PEG 8K, 25 mM sodium/potassium phosophate pH 7.0, 1 mM nicotinic acid mononucleotide, and 1 mM adenine for 30 minutes. Crystals were then soaked in the same solution, but with 30% PEG 8K instead of 20% ethylene glycol and 20% PEG 8K |