2OXM

Crystal structure of a UNG2/modified DNA complex that represent a stabilized short-lived extrahelical state in ezymatic DNA base flipping

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU300
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2006-05-11
Detector	RIGAKU RAXIS IV
Wavelength(s)	1.5418
Spacegroup name	P 21 21 21
Unit cell lengths	48.467, 65.567, 98.472
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.500 - 2.500
R-factor	0.257
Rwork	0.254
R-free	0.32800
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1emh
RMSD bond length	0.009
RMSD bond angle	1.712
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	REFMAC (5.2.0005)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	40.000	2.590
High resolution limit [Å]	2.500	2.500
Rmerge		0.599
Number of reflections	11434
<I/σ(I)>	8.3	1.91
Completeness [%]	97.1	89.4
Redundancy	4.1	3.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	293	A solution of human UNG2 (22 mg/ml) in 50 mM Tris-OAc buffer pH 7.0, 150 mM NaCl and 1mM DTT was mixed with T/M duplex DNA (2.5 mM)m incubate at room Temperature for 30 min. and then centrifugate at 10000 g for 5 min.Co-crystallizarion condition were 22% PEG 4000, 100 MM HEPES pH 6.5 and 1mM DTT, VAPOR DIFFUSION, HANGING DROP, temperature 293K

Crystallization Reagents

ID	crystal ID	solution ID	reagent name	concentration	details
1	1	1	Tris-OAc
2	1	1	NaCl
3	1	1	DTT
4	1	1	PEG 4000
5	1	1	HEPES
6	1	1	H2O
7	1	2	PEG 4000
8	1	2	HEPES
9	1	2	DTT