4M10

Crystal Structure of Murine Cyclooxygenase-2 Complex with Isoxicam

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	CCD
Collection date	2012-11-19
Detector	ADSC QUANTUM 270
Wavelength(s)	0.9792
Spacegroup name	P 2 21 21
Unit cell lengths	122.612, 134.077, 180.439
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	49.273 - 2.010
R-factor	0.1901
Rwork	0.189
R-free	0.22010
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3nt1 chain A
RMSD bond length	0.011
RMSD bond angle	1.222
Data scaling software	XSCALE
Phasing software	PHASES
Refinement software	PHENIX ((phenix.refine: 1.8.2_1309))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.090	2.120
High resolution limit [Å]	2.000	2.000
Rmerge		0.754
Number of reflections	197072
<I/σ(I)>	25.96	2.4
Completeness [%]	99.8	99.7
Redundancy	6.7	6.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	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