6BL3

Crystal Complex of Cyclooxygenase-2 with indomethacin-butyldiamine-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	2017-10-19
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.97918
Spacegroup name	C 1 2 1
Unit cell lengths	215.474, 121.776, 134.800
Unit cell angles	90.00, 123.48, 90.00

Refinement procedure

Resolution	112.436 - 2.217
R-factor	0.2054
Rwork	0.205
R-free	0.22870
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3nt1
RMSD bond length	0.003
RMSD bond angle	0.783
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX ((1.12_2829: ???))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	103.300	2.296
High resolution limit [Å]	2.217	2.217
Rmerge	0.153	1.424
Rmeas	0.183
Rpim	0.098	0.906
Number of reflections	141148	13090
<I/σ(I)>	5.97
Completeness [%]	97.8
Redundancy	3.4
CC(1/2)	0.989	0.458

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	EVAPORATION	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