2Q2T

Structure of Chlorella virus DNA ligase-adenylate bound to a 5' phosphorylated nick

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 24-ID-C
Synchrotron site	APS
Beamline	24-ID-C
Temperature [K]	100
Detector technology	CCD
Collection date	2006-03-22
Detector	ADSC QUANTUM 315
Wavelength(s)	0.9792
Spacegroup name	P 21 21 21
Unit cell lengths	66.301, 81.290, 96.418
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	38.990 - 2.300
Rwork	0.214
R-free	0.26100
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1fvi
RMSD bond length	0.006
RMSD bond angle	1.200
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.380
High resolution limit [Å]	2.300	2.300
Rmerge	0.084	0.332
Number of reflections	23105
<I/σ(I)>	14.2	3.2
Completeness [%]	96.9	95.7
Redundancy	3.6	3.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.5	295	A mixture of ChVLig (230 microM), nicked duplex DNA (220 microM) and 2 mM EDTA was added to an equal volume of a well solution containing 100 mM Bis-Tris-HCl (pH 6.5), 30 mM ammonium acetate, 22% PEG-4000. Crystals were grown at 22 C by the sitting-drop vapor diffusion method. Crystals appeared after 3 days. The crystals were transferred to buffer containing 100 mM Bis-Tris-HCl (pH 6.5), 30 mM ammonium acetate, 22% PEG-4000, 15% glycerol prior to flash-freezing in liquid nitrogen. , VAPOR DIFFUSION, SITTING DROP, temperature 295K

Crystallization Reagents

ID	crystal ID	solution ID	reagent name	concentration	details
1	1	1	Bis-Tris-HCL
10	1	3	PEG-4000
11	1	3	glycerol
2	1	1	ammonium acetate
3	1	1	PEG-4000
4	1	1	EDTA
5	1	2	PEG-4000
6	1	2	EDTA
7	1	2	ammonium acetate
8	1	3	ammonium acetate
9	1	3	EDTA