5O4C
From macrocrystals to microcrystals: a strategy for membrane protein serial crystallography
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | FREE ELECTRON LASER |
Source details | SLAC LCLS BEAMLINE CXI |
Synchrotron site | SLAC LCLS |
Beamline | CXI |
Temperature [K] | 293 |
Detector technology | PIXEL |
Collection date | 2015-04-15 |
Detector | CS-PAD CXI-1 |
Wavelength(s) | 1.89 |
Spacegroup name | P 43 21 2 |
Unit cell lengths | 226.400, 226.400, 113.700 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 36.900 - 2.800 |
R-factor | 0.16665 |
Rwork | 0.165 |
R-free | 0.19612 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 5nj4 |
RMSD bond length | 0.015 |
RMSD bond angle | 2.052 |
Data reduction software | CrystFEL (0.6.2) |
Data scaling software | CrystFEL (0.6.2) |
Phasing software | PHASER |
Refinement software | REFMAC (5.8.0158) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 36.900 | 2.890 |
High resolution limit [Å] | 2.800 | 2.800 |
Number of reflections | 69325 | |
<I/σ(I)> | 10.4 | 0.88 |
Completeness [%] | 100.0 | 100 |
Redundancy | 1342 | 361 |
CC(1/2) | 0.997 | 0.408 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 6.8 | 277 | Crystals grown from one round of seeding. Macrocrystal growth: 25 ul sitting drop. 10 ul 10 mg/ml protein, 15 ul precipitate solution Precipitate solution: 3.6 M Ammonium sulphate, 20 mM sodium phosphate buffer pH 6.8. 6 % w:v heptane-1-2-3-triol. 1 ml 2 M Ammonium sulphate reservoir Growth at 277 K for 4 days. Macrocrystals crushed into seed stock Micro-crystals 17.5 ul sitting drop. 10 ul 8.5 mg/ml protein, 7.5 ul precipitate solution 1 ml 2 M Ammonium sulphate solution |