8AAV

Human heavy chain ferritin with introduced Cys residues modified with C10 ligand

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, DESY BEAMLINE P11
Synchrotron site	PETRA III, DESY
Beamline	P11
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-08-14
Detector	DECTRIS EIGER R 4M
Wavelength(s)	1.0332
Spacegroup name	P 2 3
Unit cell lengths	181.440, 181.440, 181.440
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.540 - 2.000
R-factor	0.1449
Rwork	0.144
R-free	0.17700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5jkk
RMSD bond length	0.012
RMSD bond angle	1.675
Data reduction software	XDS
Data scaling software	Aimless (0.7.7)
Phasing software	PHASER
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	48.540	48.410	2.030
High resolution limit [Å]	2.000	10.960	2.000
Rmerge	0.052	0.039	0.181
Rmeas	0.054	0.040	0.186
Rpim	0.012	0.009	0.041
Total number of observations		16598	131455
Number of reflections	132696	891	6497
<I/σ(I)>	43.2	61.3	16
Completeness [%]	99.9	99	98.5
Redundancy	20.9	18.6	20.2
CC(1/2)	1.000	0.999	0.995

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		298	Crystallization of little amounts of protein or functionalized protein variants were performed via hanging drop vapor diffusion techniques. Reservoir solution (100 mM Tris, 500 mM MgOAc, pH 8.5) was prepared in a 24- well manual plate set. Drops were prepared on siliconized glass cover slides (Jena Bioscience) by mixing 2 microL reservoir solutions with 1 microL 50 mM Tris, 1 M NaCl, pH 7.5 buffer and 1 microL of respective ferritin variant. Plates were incubated at 298K. After one day first crystals were visible.