9EPA

Crystal structure of DARPin NY_1

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I24
Synchrotron site	Diamond
Beamline	I24
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-07-06
Detector	DECTRIS PILATUS3 6M
Wavelength(s)	0.96880
Spacegroup name	C 1 2 1
Unit cell lengths	106.558, 43.284, 32.409
Unit cell angles	90.00, 93.78, 90.00

Refinement procedure

Resolution	28.480 - 1.760
R-factor	0.1864
Rwork	0.183
R-free	0.21540
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	0.923
Data reduction software	DIALS
Data scaling software	DIALS
Phasing software	PHASER
Refinement software	PHENIX (1.19_4092)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	28.480	1.824
High resolution limit [Å]	1.760	1.761
Number of reflections	14698	14698
<I/σ(I)>	6.53
Completeness [%]	99.5
Redundancy	6.5
CC(1/2)	0.982	0.796

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		298	NY_1/HLA-A0201/NY-ESO1157-165(9V) complex was concentrated to 10 mg/mL in 20 mM HEPES, 10% glycerol, 300 mM NaCl pH 7.5. Crystallization was performed within the protein science facility at Karolinska Institutet (https://ki.se/en/mbb/psf-mx). Crystals of NY_1 alone were obtained in condition 1-48 (pH 8.5; 0.12 M alcohol mix; 0.1 M buffer system 3; 37.5% (v/v) MPD_P1K_P3350) of the Morpheus screen (Molecular Dimensions Ltd.). For condition screening, the Mosquito robot was used to setup 1:2, 1:1 and 2:1 drop ratios in 96-well sitting-drop iQ TTP Labtech plates (TTP Labtech).