8EK4

De novo designed ice-binding proteins from twist-constrained helices

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-03-14
Detector	DECTRIS EIGER2 X 16M
Wavelength(s)	0.97918
Spacegroup name	P 21 21 21
Unit cell lengths	54.582, 67.427, 74.326
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	49.940 - 2.350
R-factor	0.2614
Rwork	0.259
R-free	0.31370
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Designed model
RMSD bond length	0.002
RMSD bond angle	0.366
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX (1.17.1_3660)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	59.940	2.430
High resolution limit [Å]	2.350	2.350
Rmerge	0.114	1.379
Rmeas	0.119
Rpim	0.033	0.381
Number of reflections	11928	1172
<I/σ(I)>	13.8	1.55
Completeness [%]	99.6
Redundancy	13.2
CC(1/2)	0.999	0.753

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	0.02M 1,6-hexanediol, 0.02M 1-butanol, 0.02M 1,2-propanediol, 0.02M 2-propanol, 0.02M 2-propanol, 0.02M 1,4-butanediol, 0.02M 1,3-propanediol, 0.0466M pH 8.5 Tris (base), 0.0534M pH 8.5 Bicine, 20% v/v PEG 500 MME, and 10% w/v PEG 20,000