6B8G
Twice-Contracted Human Heavy-Chain Ferritin Crystal-Hydrogel Hybrid
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | SSRL BEAMLINE BL9-2 |
Synchrotron site | SSRL |
Beamline | BL9-2 |
Temperature [K] | 100 |
Detector technology | PIXEL |
Collection date | 2017-07-22 |
Detector | DECTRIS PILATUS 6M |
Wavelength(s) | 0.97946 |
Spacegroup name | F 4 3 2 |
Unit cell lengths | 179.950, 179.950, 179.950 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 34.631 - 1.130 |
R-factor | 0.1036 |
Rwork | 0.102 |
R-free | 0.12130 |
RMSD bond length | 0.014 |
RMSD bond angle | 1.370 |
Data reduction software | iMOSFLM |
Data scaling software | Aimless |
Phasing software | PHASER |
Refinement software | PHENIX (1.12_2829) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 63.620 | 1.150 |
High resolution limit [Å] | 1.130 | 1.130 |
Rmerge | 0.131 | 1.131 |
Number of reflections | 92912 | 4518 |
<I/σ(I)> | 19.1 | 2.3 |
Completeness [%] | 100.0 | 100 |
Redundancy | 32.2 | 13.8 |
CC(1/2) | 0.999 | 0.628 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 293 | Reservoir: 500 uL of 25 mM HEPES pH 7.0 with 10 mM CaCl2 Well: 2 uL of reservoir solution and 2 uL of 2.5 uM ferritin (by 24-mer) in 15 mM TRIS pH 7.4 with 150 mM NaCl After crystals formed, a crystal was harvested and soaked in a buffered solution comprised of 25 mM HEPES pH 7.0, 30 mM CaCl2, 8.6 % (w/v) sodium acrylate, 2.5 % (w/v) acrylamide, and 0.2 % (w/v) Bis-acrylamide overnight. Crystal was transferred to a solution containing 1 % APS and 1% TEMED with 4 M NaCl for 5 min. Crystal was then transferred to a clean slide and soaked in 30 uL H2O for 5 min. The H2O was removed the crystal was soaked in 4 M NaCl for 2 min. The crystal was removed, and soaked in 30 uL H2O for 5 min. This solution was removed and the crystal was soaked in 1 M CaCl2 for 2 min. The crystal was removed, cryoprotected in perfluoro polyether, and frozen in liquid N2. |