5IZZ
Crystal structure of a marine metagenome TRAP solute binding protein specific for aromatic acid ligands (Sorcerer II Global Ocean Sampling Expedition, unidentified microbe, locus tag GOS_1523157, Triple Surface Mutant K158A_K223A_K313A) in complex with metahydroxyphenylacetate, thermal exchange of ligand
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | ROTATING ANODE |
Source details | RIGAKU MICROMAX-007 HF |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2016-03-15 |
Detector | DECTRIS PILATUS 200K |
Wavelength(s) | 1.54056 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 39.784, 84.275, 45.910 |
Unit cell angles | 90.00, 113.48, 90.00 |
Refinement procedure
Resolution | 29.786 - 1.600 |
R-factor | 0.1166 |
Rwork | 0.115 |
R-free | 0.15310 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5i7i |
RMSD bond length | 0.008 |
RMSD bond angle | 0.990 |
Data reduction software | HKL-3000 |
Data scaling software | HKL-2000 |
Phasing software | MOLREP |
Refinement software | PHENIX ((1.10.1_2155)) |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 100.000 | 100.000 | 1.630 |
High resolution limit [Å] | 1.600 | 4.340 | 1.600 |
Rmerge | 0.052 | 0.035 | 0.265 |
Rmeas | 0.059 | 0.039 | 0.322 |
Rpim | 0.027 | 0.015 | 0.180 |
Total number of observations | 143046 | ||
Number of reflections | 34550 | ||
<I/σ(I)> | 12.3 | ||
Completeness [%] | 94.0 | 99.8 | 80.3 |
Redundancy | 4.1 | 6.2 | 2.8 |
CC(1/2) | 0.999 | 0.866 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 8.5 | 298 | Protein (10 mM HEPES pH 7.5, 5 mM DTT, 10 mM 3-Hydroxyphenylacetic acid, soak 30 minutes at 60 C); Reservoir (MCSG1 (A2), 0.1 M CHES pH 9.5, 30 %(w/v) PEG 3000); Cryoprotection (20% diethylene glycol, 80% reservoir) |