4Z7H

Crystal structure of human IRE1 cytoplasmic kinase-RNase region - complex with imidazopyridine compound 3

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04-1
Synchrotron site	Diamond
Beamline	I04-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2013-09-24
Detector	DECTRIS PILATUS 2M
Wavelength(s)	0.92
Spacegroup name	P 21 21 21
Unit cell lengths	78.678, 81.957, 168.682
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	46.360 - 2.900
R-factor	0.2015
Rwork	0.200
R-free	0.22480
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4z7g
RMSD bond length	0.010
RMSD bond angle	1.140
Data reduction software	XDS
Data scaling software	Aimless
Refinement software	BUSTER (2.10.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	46.360	3.080
High resolution limit [Å]	2.900	2.900
Rmerge	0.201	1.440
Number of reflections	24770
<I/σ(I)>	7.5	1.4
Completeness [%]	99.7	99
Redundancy	6.6	6.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		291	Apo-hIRE1 was crystallized by hanging-drop vapor diffusion by mixing IRE1 (at 2 mg/mL) with reservoir solution containing 18-20% PEG3350, 0.2 M sodium malonate and 0.1 Bis-Trispropane (pH 6.5) in a 1:1 ratio. Crystals were briefly (30 sec) soaked in 22% PEG3350, 0.2 M Na Malonate, 0.1 M Bis-Trispropane (pH 7.5) and 25% ethylene glycol before plunging into liquid N2.