2OUU

crystal structure of PDE10A2 mutant D674A in complex with cGMP

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	2006-04-27
Detector	ADSC QUANTUM 315
Spacegroup name	P 21 21 21
Unit cell lengths	48.717, 81.994, 153.977
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	30.000 - 1.520
Rwork	0.204
R-free	0.22200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	pde10a2 native
RMSD bond length	0.006
RMSD bond angle	1.300
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	AMoRE
Refinement software	CNS (1.1)

Data quality characteristics

	Overall
Low resolution limit [Å]	30.000
High resolution limit [Å]	1.520
Rmerge	0.082
Number of reflections	95665
<I/σ(I)>	10
Completeness [%]	99.9
Redundancy	9.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	277	0.1 M HEPES, pH 7.5, 0.1 M MgCl2, 100 mM BME, and 13% PEG3350. The D674A mutant crystals were soaked in 30 mM cGMP in a buffer of 16% PEG8000, 0.1 M HEPES, pH 7.5, 0.1 M MgCl2, 60 mM BME for 1.5 hours, VAPOR DIFFUSION, HANGING DROP, temperature 277K