2NO3

Novel 4-anilinopyrimidines as potent JNK1 Inhibitors

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU300
Temperature [K]	110
Detector technology	CCD
Collection date	2005-01-06
Detector	MAR CCD 165 mm
Wavelength(s)	1.5418
Spacegroup name	P 32 2 1
Unit cell lengths	157.201, 157.201, 123.362
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	19.980 - 3.200
R-factor	0.2332
Rwork	0.206
R-free	0.27600
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	entry 2h96
RMSD bond length	0.008
RMSD bond angle	1.400
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	CNX
Refinement software	CNX (2002)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	3.310
High resolution limit [Å]	3.200	3.200
Rmerge	0.083	0.454
Number of reflections	27814
<I/σ(I)>	15.3	1.6
Completeness [%]	94.5	98.3
Redundancy	5.6	4.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	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 2 UL WELL SOLUTION. WELL SOLUTION:2.8-3.1 M AMMONIUM SULFATE, 10- 14% GLYCEROL. FOR CO-CRYSTALLIZATION EXPERIMENT WITH THE COMPOUND, THE 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 6.20, VAPOR DIFFUSION, temperature 277.0K