5T7E
Crystal structure of Streptomyces hygroscopicus Bialaphos Resistance (BAR) protein in complex with Coenzyme A and L-phosphinothricin
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | APS BEAMLINE 24-ID-C |
Synchrotron site | APS |
Beamline | 24-ID-C |
Temperature [K] | 90 |
Detector technology | PIXEL |
Collection date | 2015-08-08 |
Detector | DECTRIS PILATUS 6M-F |
Wavelength(s) | 0.987 |
Spacegroup name | P 1 21 1 |
Unit cell lengths | 64.180, 66.110, 86.150 |
Unit cell angles | 90.00, 103.06, 90.00 |
Refinement procedure
Resolution | 62.520 - 1.800 |
R-factor | 0.2098 |
Rwork | 0.208 |
R-free | 0.24710 |
Structure solution method | MOLECULAR REPLACEMENT |
RMSD bond length | 0.002 |
RMSD bond angle | 0.590 |
Data reduction software | iMOSFLM |
Data scaling software | SCALA |
Phasing software | PHASER |
Refinement software | PHENIX ((dev_2499: ???)) |
Data quality characteristics
Overall | |
Low resolution limit [Å] | 62.520 |
High resolution limit [Å] | 1.800 |
Number of reflections | 93996 |
<I/σ(I)> | 5.41 |
Completeness [%] | 83.6 |
Redundancy | 1.7 |
CC(1/2) | 0.960 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 293 | BAR protein was incubated with 1 mM acetyl-CoA for >2 hour prior to setting crystal trays. Crystals of BAR were obtained after 3 days at 20C in hanging drops containing 1 uL of protein solution (7.5 mg/mL) and 1 uL of reservoir solution (0.18 M calcium acetate, 0.1 M Tris-HCl pH 7, 18% (w/v) PEG 3000, 0.2% (v/v) N-nonyl Beta-D-glucopyranoside, 1 mM acetyl-CoA). Several crystals were soaked in reservoir solution supplemented with 30 mM L-phosphinothricin for 30-60 min before freezing. Crystals were frozen in reservoir solution supplemented with 15% (v/v) ethylene glycol. Acetylation of phosphinothricin occurred during soaking as no density for the acetyl group of acetyl-CoA was observed in the BAR/CoA/phosphinothricin ternary complex. |