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.