6BL4

Crystal Complex of Cyclooxygenase-2 with indomethacin-ethylenediamine-dansyl conjugate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-06-24
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.97918
Spacegroup name	C 1 2 1
Unit cell lengths	214.795, 120.414, 134.417
Unit cell angles	90.00, 123.60, 90.00

Refinement procedure

Resolution	111.959 - 2.220
R-factor	0.1895
Rwork	0.189
R-free	0.21950
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3nt1
RMSD bond length	0.012
RMSD bond angle	1.013
Data reduction software	XDS
Data scaling software	Aimless (0.1.27)
Phasing software	PHASER
Refinement software	PHENIX (1.12_2829)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	111.960	111.960	2.340
High resolution limit [Å]	2.220	7.020	2.220
Rmerge	0.110	0.027	0.677
Rmeas	0.144	0.036	0.886
Rpim	0.093	0.024	0.566
Total number of observations	325121
Number of reflections	138444	4542	20005
<I/σ(I)>	9.3
Completeness [%]	98.7	99.3	97.9
Redundancy	2.3	2.3	2.4
CC(1/2)	0.985	0.997	0.555

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.7	291	mCOX-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 uL of the protein-inhibitor complex with 3 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, VAPOR DIFFUSION, HANGING DROP, temperature 291K