8F0L

Crystal Structure of the Human T cell Receptor CD3(EPSILON) N-Terminal Peptide Complexed with ADI-26906 FAB

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	MAX IV BEAMLINE BioMAX
Synchrotron site	MAX IV
Beamline	BioMAX
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-04-14
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.9763
Spacegroup name	P 1 21 1
Unit cell lengths	73.634, 53.084, 113.934
Unit cell angles	90.00, 99.82, 90.00

Refinement procedure

Resolution	48.600 - 1.810
Rwork	0.166
R-free	0.19500
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4irz
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (v1.19.2)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	66.310	1.841
High resolution limit [Å]	1.810	1.810
Rmerge	0.074	0.872
Rmeas	0.080	0.942
Rpim	0.030	0.352
Number of reflections	79385	3938
<I/σ(I)>	13.8	2.1
Completeness [%]	99.9	99.6
Redundancy	6.9	7
CC(1/2)	0.998	0.746

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	293	Crystals of the ADI-26906 Fab complexed to CD3 epsilon N13 peptide were obtained by the sitting drop vapor diffusion method. The ADI-26906 Fab buffered in PBS, was first buffer-exchanged into 20 mM Bis-Tris pH 6.0 and 150 mM NaCl. The Fab (0.31 mM) was mixed with the N13 peptide (7.8 mM) at a Fab:peptide ratio of 1:25 and incubated on ice for 1.5 h. A BCS screen (Molecular Dimensions Ltd.) was set up using a mosquito crystallization robot (STP Labtech), with each drop consisting of 100 nL protein and 100 nL of reservoir solution, and left to equilibrate against a 40 uL reservoir solution at 293 K. After a few days, crystals were obtained in condition A6 (0.1 M MES pH 6.5, 25% w/v PEG Smear Broad).