4XI2

Crystal Structure of an auto-inhibited form of Bruton's Tryrosine Kinase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	CHESS BEAMLINE A1
Synchrotron site	CHESS
Beamline	A1
Temperature [K]	100
Detector technology	CCD
Collection date	2002-11-02
Detector	ADSC QUANTUM 210
Wavelength(s)	0.9474
Spacegroup name	P 31 2 1
Unit cell lengths	132.202, 132.202, 107.632
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	43.273 - 2.600
R-factor	0.2306
Rwork	0.230
R-free	0.24830
RMSD bond length	0.007
RMSD bond angle	1.114
Data reduction software	DENZO (1.97.8)
Data scaling software	SCALEPACK (1.97.7)
Phasing software	SHARP (1.4)
Refinement software	PHENIX ((phenix.refine: dev_1839))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	43.300	2.660
High resolution limit [Å]	2.600	2.600
Number of reflections	30976
<I/σ(I)>	10.6	1.1
Completeness [%]	91.9	69.6
Redundancy	4.8	3.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5	277	Mix purified dimer protein at 5 mg/ml in 25 mM TRIS.Cl, pH 8.5, 10 mM NaCl, 5 mM DTT with an equal volume of the precipitant solution, 100 mM TRIS.Cl, pH 8.5, 200 mM Na.acetate, 7.5 % PEG 4,000, then suspend over 1 ml of precipitant solution. Wedge shaped crystals grew to approximately 0.1 mm on a side after a period of two weeks. Crystals were frozen by first transferring them, in four sequential steps, to a solution of 100 mM TRIS.Cl, pH 8.5, 200 mM Na.acetate, 15 % PEG 4,000. This was followed by transfer in six sequential steps to a solution containing 100 mM TRIS.Cl, pH 8.5, 200 mM Na.acetate, 15 % PEG 4,000, 30% glucose