2NUN

The structure of the type III effector AvrB complexed with ADP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 22-ID
Synchrotron site	APS
Beamline	22-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2005-10-30
Detector	MAR CCD 165 mm
Wavelength(s)	1.0000
Spacegroup name	P 65
Unit cell lengths	122.717, 122.717, 64.091
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	20.000 - 2.400
R-factor	0.232
Rwork	0.232
R-free	0.25400
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1nh1
RMSD bond length	0.007
RMSD bond angle	1.300
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.230
High resolution limit [Å]	2.150	2.150
Rmerge	0.093	0.318
Number of reflections	24091
<I/σ(I)>	28	2.63
Completeness [%]	80.4	32
Redundancy	4.9	3.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	9	277	Protein (8-10 mg/ml) was mixed with well solution (100 mM Glycine, pH 9.0, 20-30% PEG 550 MME)at a 1:1 ratio. Microseeding was employed to obtain better crystals. Nucleotides were incorporated by exchanging drop and resevoir solutions with 20 l 27% PEG 500 MME and 100 mM Tris 7.5 plus 5 mM MgCl2, followed by a final exchange of this solution plus 5 mM nucleotide. Nucleotides were soaked for ~ 1 day. , VAPOR DIFFUSION, SITTING DROP, temperature 277K