3QI4

Crystal structure of PDE9A(Q453E) in complex with IBMX

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
Detector	ADSC QUANTUM 315
Wavelength(s)	1.1
Spacegroup name	P 41 21 2
Unit cell lengths	103.409, 103.409, 269.950
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	30.000 - 2.500
R-factor	0.213
Rwork	0.213
R-free	0.23900
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	1.200
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS (1.1)

Data quality characteristics

	Overall
Low resolution limit [Å]	30.000
High resolution limit [Å]	2.500
Rmerge	0.094
Number of reflections	50388
<I/σ(I)>	12.2
Completeness [%]	97.6
Redundancy	13.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	277	The catalytic domain of the PDE9Q453E mutant (10-15 mg/mL, amino acids 181-506) was stored in a buffer of 50 mM NaCl, 20 mM Tris.HCl pH 7.5, 1 mM beta-mercaptoethanol, and 1 mM EDTA. After mixing with 2 mM IBMX, the PDE9Q453E-IBMX complex was crystallized by hanging drop vapor diffusion against the well buffer of 2.0 M Na formate, 0.1 M HEPES pH 7.5, 5% xylitol at 277K., VAPOR DIFFUSION, HANGING DROP