5OHJ

Human phosphodiesterase 4B catalytic domain in complex with a pyrrolidinyl inhibitor.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04-1
Synchrotron site	Diamond
Beamline	I04-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2012-10-31
Detector	DECTRIS PILATUS 2M
Wavelength(s)	0.92
Spacegroup name	P 21 21 21
Unit cell lengths	55.270, 55.730, 225.560
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.610 - 1.600
R-factor	0.18549
Rwork	0.184
R-free	0.21153
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	dimer of the catalytic domain of PDE4D
RMSD bond length	0.019
RMSD bond angle	1.880
Data reduction software	XDS
Data scaling software	XDS
Phasing software	PHASER
Refinement software	REFMAC (5.7.0029)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	29.610	1.550
High resolution limit [Å]	1.510	1.510
Rmerge	0.046	0.467
Rpim	0.041	0.149
Number of reflections	85681	2246
<I/σ(I)>	22.4	2.6
Completeness [%]	77.7	28.3
Redundancy	6.4	3.7
CC(1/2)	0.999	0.791

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.5	293	Drops comprised 2microliters protein solution mixed with reservoir buffer consisting of 100mM sodium acetate pH 4.5, 14% PEG400 and 50mM calcium acetate. After 2 days they were streak seeded with crystals obtained from rolipram complex seed stock. Crystals, up to 100 microns in largest dimension, grew from precipitate after 1-2 weeks. Crystals were transferred to a cryoprotectant solution (100mM sodium acetate pH 4.5, 14% PEG400, 50mM calcium acetate and 20% glycerol) then flash-frozen prior to X-ray data collection.