3N8M

Crystal Structure of the Grb2 SH2 Domain in Complex with An Acyclic Ligand Having the Sequence pYVNVP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2007-07-27
Detector	MAR scanner 345 mm plate
Wavelength(s)	1.5418
Spacegroup name	P 41 21 2
Unit cell lengths	49.240, 49.240, 86.128
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	34.820 - 2.000
Rwork	0.187
R-free	0.22390
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2huw
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	MOLREP
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	34.820	1.930
High resolution limit [Å]	1.860	1.860
Rmerge	0.066	0.152
Number of reflections	9451
<I/σ(I)>	60.6	21.1
Completeness [%]	99.6	97
Redundancy	22.1	19.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.5	298	Ligand in lyophilized powder form was dissolved in a 7.2 mg/mL solution of Grb2 SH2 in water such to give a protein/ligand molar ratio of 1:1.7. 4 uL of this solution was mixed with 3 uL of 12% v/v glycerol, 1.5 M ammonium sulfate, 0.1 M TRIS, pH 8.5 to create the hanging drop, which yielded crystals of the protein-ligand complex after 8 weeks, VAPOR DIFFUSION, HANGING DROP, temperature 298K