5W58

Crystal Complex of Cyclooxygenase-2: (S)-ARN-2508 (a dual COX and FAAH inhibitor)

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	CCD
Collection date	2016-03-02
Detector	ADSC QUANTUM 315
Wavelength(s)	0.97918
Spacegroup name	I 41 2 2
Unit cell lengths	173.774, 173.774, 203.304
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	66.048 - 2.267
R-factor	0.1697
Rwork	0.169
R-free	0.18860
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3nt1
RMSD bond length	0.021
RMSD bond angle	1.270
Data reduction software	XDS
Data scaling software	Aimless (0.5.21)
Phasing software	PHASER
Refinement software	PHENIX ((1.10.1_2155: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	132.100	2.320
High resolution limit [Å]	2.267	2.270
Rmerge	0.155
Rmeas	0.161	4.323
Rpim	0.042	1.136
Number of reflections	71762
<I/σ(I)>	19.4
Completeness [%]	99.8	96.5
Redundancy	14.6	14.2
CC(1/2)	0.999	0.311

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	291	COX-2 protein reconstituted with a 2-fold molar excess of heme in phosphtate buffer, pH 6.7, 100 mM NaCl, 1.2% (w/v) -OG, and 0.1% NaN3, and 10-fold molar excess of inhibitors from 25 mM DMSO stocks were added to protein samples. Mixing 3.5 uL of the protein-inhibitor complex with 3.5 uL crystallization solution containing 50 mM EPPS, pH 8.0, 120 mM MgCl2, 22-26% PEG MME-550 against reservoir solutions comprised of 50 mM EPPS pH 8.0, 120 mM MgCl2, 22-26% PEG MME-550