4M11

Crystal Structure of Murine Cyclooxygenase-2 Complex with Meloxicam

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 315
Wavelength(s)	0.9792
Spacegroup name	P 2 21 21
Unit cell lengths	121.627, 133.538, 180.232
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	49.954 - 2.450
R-factor	0.2032
Rwork	0.202
R-free	0.23060
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3nt1 chain A
RMSD bond length	0.009
RMSD bond angle	1.166
Data scaling software	SCALA (3.3.20)
Phasing software	PHASES
Refinement software	PHENIX (1.8.2_1309)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	49.954	49.954	2.580
High resolution limit [Å]	2.450	7.740	2.450
Rmerge		0.044	0.654
Total number of observations		21384	87103
Number of reflections	106951
<I/σ(I)>	9.2	11	1.1
Completeness [%]	98.9	96.4	98.4
Redundancy	6.1	6	5.7

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