9BD2

MAGE-A3 MHD crystal soaked with KL861

This is a non-PDB format compatible entry.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-E
Synchrotron site	APS
Beamline	24-ID-E
Temperature [K]	100
Detector technology	PIXEL
Collection date	2022-07-29
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.9792
Spacegroup name	C 2 2 21
Unit cell lengths	55.659, 103.402, 325.668
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	81.420 - 2.240
R-factor	0.2202
Rwork	0.218
R-free	0.26020
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.004
RMSD bond angle	0.717
Data reduction software	DIALS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.20.1_4487)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	81.420	2.270
High resolution limit [Å]	2.230	2.230
Number of reflections	46101	2251
<I/σ(I)>	11.4
Completeness [%]	99.9
Redundancy	8.9
CC(1/2)	0.990	0.321

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		277	MAGE-A3 MHD was buffer-exchanged into 25 mM HEPES, 150 mM NaCl, pH 7.5 buffer using 10 kDa MWCO Centricon and concentrated to 5 mg/mL. MAGE-A3 MHD was crystallized using the hanging drop vapor diffusion method in 16% PEG8000, 40 mM potassium phosphate, 20% glycerol. For each hanging drop, 2 ul of protein solution was mixed with an equal volume of well solution and incubated at 4 C. Once the crystals reached their maximum size, they were transferred to a cryo-protectant solution (well solution containing 30% glycerol and 1 mM KL861) and soaked with KL861 for 3.5 hours. The soaked crystals were flash-frozen in liquid nitrogen.