6B8F

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	2017-04-01
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.97946
Spacegroup name	F 4 3 2
Unit cell lengths	180.040, 180.040, 180.040
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.010 - 1.060
R-factor	0.0916
Rwork	0.091
R-free	0.10260
RMSD bond length	0.013
RMSD bond angle	1.317
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]	63.650	1.080
High resolution limit [Å]	1.060	1.060
Rmerge	0.060	0.222
Number of reflections	111189	4683
<I/σ(I)>	24.8	3.2
Completeness [%]	99.0	86.4
Redundancy	14	2.4
CC(1/2)	0.999	0.897

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. 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.