3K3E

Crystal structure of the PDE9A catalytic domain in complex with (R)-BAY73-6691

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X29A
Synchrotron site	NSLS
Beamline	X29A
Temperature [K]	100
Detector technology	CCD
Collection date	2009-06-05
Detector	ADSC QUANTUM 315
Wavelength(s)	1.1000
Spacegroup name	P 41 21 2
Unit cell lengths	103.190, 103.190, 270.849
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	30.000 - 2.700
R-factor	0.23
Rwork	0.230
R-free	0.25800
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	1.200
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	AMoRE
Refinement software	CNS (1.2)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.800
High resolution limit [Å]	2.700	2.700
Rmerge	0.084	0.590
Number of reflections	41185
<I/σ(I)>	12.2	100
Completeness [%]	100.0	100
Redundancy	11

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	277	soaking PDE9A2-IBMX crystal in the buffer of 0.1 M HEPES pH 7.5, 3.6 M sodium formate, and 2 mM R6691 at 25 deg C for 3 days. The PDE9A2-IBMX crystals were grown by (1) mixing 10-15 mg/mL PDE9A2 catalytic domain (amino acids 181-506) with 2 mM IBMX in a buffer of 50 mM NaCl, 20 mM Tris-HCl pH 7.5, 1 mM beta-mercaptoethanol, 1 mM EDTA, and (2) the vapor diffusion (hanging drop) at 4 deg C. The protein drops contained 2 microliters PDE9A2-IBMX complex and 2 microliters well solution of 0.1 M HEPES pH 7.5 and 3.0 M sodium formate, VAPOR DIFFUSION, HANGING DROP, temperature 277K