2G01

Pyrazoloquinolones as Novel, Selective JNK1 inhibitors

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 17-ID
Synchrotron site	APS
Beamline	17-ID
Temperature [K]	110
Detector technology	CCD
Collection date	2004-10-18
Detector	ADSC QUANTUM 4
Spacegroup name	P 32 2 1
Unit cell lengths	150.622, 150.622, 118.996
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	19.890 - 3.500
R-factor	0.286
Rwork	0.283
R-free	0.35400
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1jnk JNK3 KINASE
RMSD bond length	0.008
RMSD bond angle	1.400
Data reduction software	SDMS
Data scaling software	HKL-2000
Phasing software	CNX
Refinement software	CNX (2002)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	3.620
High resolution limit [Å]	3.500	3.500
Rmerge	0.065	0.365
Number of reflections	15793
<I/σ(I)>	27.9	2.6
Completeness [%]	81.7	63.7
Redundancy	5.8	5.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.2	277	Protein was preincubated with the JIP1 peptide at a 5x molar excess. Protein concentration 9-12.6 mg/mL. Hanging drops consisted of 2uL protein plus 2uL well solution over 1 mL of well solution. Well solution: 2.8-3.1 M Ammonium Sulfate, 10-14% glycerol. For Co-crystallization experiment compound was dissolved in DMSO at 100 mM concentration. Allow to incubate for at least an hour on ice. Solution was spun for 5 minutes at 2000g prior to setting up for crystallization. , pH 6.2, VAPOR DIFFUSION, HANGING DROP, temperature 277.0K