6ASP

Structure of Grp94 with methyl 3-chloro-2-(2-(1-(2-ethoxybenzyl)-1 H-imidazol-2-yl)ethyl)-4,6-dihydroxybenzoate, a Grp94-selective inhibitor and promising therapeutic lead for treating myocilin-associated glaucoma

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 22-BM
Synchrotron site	APS
Beamline	22-BM
Temperature [K]	197.5
Detector technology	CCD
Collection date	2016-11-22
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	1.00000
Spacegroup name	P 21 21 21
Unit cell lengths	65.661, 84.772, 96.013
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	38.776 - 2.696
R-factor	0.1836
Rwork	0.178
R-free	0.23770
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2gfd
RMSD bond length	0.010
RMSD bond angle	1.244
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	PHASER
Refinement software	PHENIX ((1.10.1_2155: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	38.780	2.792
High resolution limit [Å]	2.696	2.696
Rmerge	0.130	0.675
Number of reflections	15293
<I/σ(I)>	22.46
Completeness [%]	99.0
Redundancy	13.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.8	289.2	1:1 30 mg/mL protein in 100 mM bicine, pH 7.8 against mother liquor (37% PEG400, 5% glycerol, 100 mM bicine, pH 7.8, 75 mM magnesium chloride). Crystals were harvested, and soaked for 48 hours in mother liquor containing 3.69 mM inhibitor. After soaking, glycerol was added to ~25% final concentration.