4ORZ

HIV-1 Nef protein in complex with single domain antibody sdAb19 and an engineered Hck SH3 domain

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X10SA
Synchrotron site	SLS
Beamline	X10SA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2012-02-10
Detector	PSI PILATUS 6M
Wavelength(s)	0.979400
Spacegroup name	P 41
Unit cell lengths	73.000, 73.000, 71.000
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	41.750 - 2.000
R-factor	0.2036
Rwork	0.202
R-free	0.23640
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3rbb
RMSD bond length	0.008
RMSD bond angle	1.288
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX ((phenix.refine: 1.8.1_1168))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	41.750	2.150
High resolution limit [Å]	2.000	2.100
Number of reflections	47918
Completeness [%]	100.0	100

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	9	293	About 0.1 micro L of protein solution at 10 mg/ml concentration was mixed with 0.1 micro L of reservoir solution from a 70 L reservoir in 96-well Hampton 3553 crystallization plates. Initial crystals of NefSF2 sdAb19 SH3B6 could be obtained in 0.2 M potassium formate and 20% polyethylene glycol (PEG) 3350. Crystal conditions were optimized to 0.2 M potassium formate, 17.5% polyethylene glycol (PEG) 3350 and 0.35 M ammonium chloride grown by hanging-drop vapor diffusion in Linbro crystallization plates. , pH 9.0, VAPOR DIFFUSION, HANGING DROP, temperature 293K