3UBY

Crystal structure of human alklyadenine DNA glycosylase in a lower and higher-affinity complex with DNA

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 12.3.1
Synchrotron site	ALS
Beamline	12.3.1
Temperature [K]	100
Detector technology	CCD
Collection date	2006-07-11
Detector	ADSC QUANTUM 315
Wavelength(s)	1.116
Spacegroup name	P 43
Unit cell lengths	41.168, 41.168, 262.546
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	65.650 - 2.000
R-factor	0.22176
Rwork	0.219
R-free	0.26545
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1bnk
RMSD bond length	0.007
RMSD bond angle	1.104
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	REFMAC (5.5.0109)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	66.000	2.053
High resolution limit [Å]	2.000	2.000
Number of reflections	26998
Completeness [%]	100.0	100

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		298	An equimolar ratio of delta79AAG and 13-mer single-stranded (ss) EDC DNA were mixed to form a protein-DNA complex concentration of 0.3 mM in the complex buffer (20 mM HEPES-NaOH, pH 7.5, 100 mM NaCl, 0.1 mM EDTA, 5% v/v glycerol and 1 mM DTT). The complex was incubated on ice for 15 min and used for crystallization. Crystals were obtained upon mixing 1 uL of protein-DNA complex and 1 uL of reservoir solution (100 mM BIS-TRIS, pH 5.5, 200 mM cesium chloride and 20% polyethylene glycol (PEG) 3350) over 0.5 ml of reservoir solution. Crystals appeared after incubation for 14 days at 22 degrees C, VAPOR DIFFUSION, HANGING DROP, temperature 298K