1PTW

The Crystal Structure of AMP-Bound PDE4 Suggests a Mechanism for Phosphodiesterase Catalysis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 14-BM-C
Synchrotron site	APS
Beamline	14-BM-C
Temperature [K]	100
Detector technology	CCD
Collection date	2002-08-08
Detector	ADSC QUANTUM 4
Wavelength(s)	0.9
Spacegroup name	P 21 21 21
Unit cell lengths	99.207, 111.229, 159.650
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	99.000 - 2.400 *
Rwork	0.229
R-free	0.27400
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.007
RMSD bond angle	1.180
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS (1.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	99.000	2.380
High resolution limit [Å]	2.400 *	2.400 *
Rmerge	0.075 *	0.450 *
Total number of observations	404282 *
Number of reflections	64052 *
<I/σ(I)>	21.9	3.9
Completeness [%]	91.8 *	87.9 *
Redundancy	6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	4 *	50 mM HEPES, 15% PEG 3350, 25% ethylene glycol, 5% methanol, 5% DMSO, pH 7.5, VAPOR DIFFUSION, HANGING DROP, temperature 277K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	reservoir	HEPES	50 (mM)	pH7.5
10	1	drop	Tris-HCl	20 (mM)
11	1	drop	beta-mercaptoethanol	1 (mM)
2	1	reservoir	PEG3350	15 (%)
3	1	reservoir	ethylene glycol	25 (%)
4	1	reservoir	methanol	5 (%)
5	1	reservoir	DMSO	5 (%)
6	1	drop	cAMP	10 (mM)
7	1	drop	zinc sulfate	0.4 (mM)
8	1	drop	protein	15 (mg/ml)
9	1	drop		50 (mM)	pH7.5