4GH6

Crystal structure of the PDE9A catalytic domain in complex with inhibitor 28

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PHOTON FACTORY BEAMLINE BL-17A
Synchrotron site	Photon Factory
Beamline	BL-17A
Temperature [K]	100
Wavelength(s)	1.0
Spacegroup name	P 41 21 2
Unit cell lengths	104.276, 104.276, 270.073
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	30.000 - 2.700
Rwork	0.212
R-free	0.24500
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	1.300
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	AMoRE
Refinement software	CNS

Data quality characteristics

	Overall
Low resolution limit [Å]	30.000
High resolution limit [Å]	2.700
Rmerge	0.093
Number of reflections	41120
<I/σ(I)>	17.4
Completeness [%]	99.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	298	PDE9A2 (15 mg/mL) in a buffer of 50 mM NaCl, 20 mM Tris.HCl pH 7.5, 1 mM beta-mercaptoethanol and 1 mM EDTA was mixed with 2 mM IBMX for 2h before setting-up of crystallization against the well buffer of 2.0 M Na formate, 0.1 M HEPES pH 7.5, 5% xylitol at 4C. The PDE9A-IBMX complex crystals were formed after 1d and reached the maximum size in 2 weeks. Crystals of the PDE9A-28 complex were prepared by soaking PDE9-IBMX co-crystals in the crystallization buffer plus 5 mM 28 at 25C for 3 days, VAPOR DIFFUSION, HANGING DROP, temperature 298K