2B3A | pdb_00002b3a

Solution structure of the Ras-binding domain of the Ral Guanosine Dissociation Stimulator

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D NOESY	20 mM potassium buffer at ph 7.0, 2 mM dithioerythritol (DTE), 0.5 mM EDTA, 0.5 mM NaN3, 0.1 mM 2,2-dimethyl-2-silapentane sulfonic acid (DSS) 1.0 mM RalGDS-RBD non labeled	95% H2O/5% D2O (v/v)	20 mM potassium buffer	7.0	ambient	298
2	3D_15N-separated_NOESY	20 mM potassium buffer at ph 7.0, 2 mM dithioerythritol (DTE), 0.5 mM EDTA, 0.5 mM NaN3, 0.1 mM 2,2-dimethyl-2-silapentane sulfonic acid (DSS) 2.9 mM RalGDS-RBD 15N/13C labeled	95% H2O/5% D2O (v/v)	20 mM potassium buffer	7.0	ambient	298
3	2D NOESY	20 mM potassium buffer at ph 7.0, 2 mM dithioerythritol (DTE), 0.5 mM EDTA, 0.5 mM NaN3, 0.1 mM 2,2-dimethyl-2-silapentane sulfonic acid (DSS) 1.0 mM RalGDS-RBD non labeled	100% D2O	20 mM potassium buffer	7.0	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DMX	800
2	Bruker	DMX	600

NMR Refinement
Method	Details	Software
simulated annealing first in torsion angle space followed by simulated annealing in cartesian space. Final refinement in explicit solvent (H2O).	The structures are based on a total of 1680 restraints, 1550 are NOE-derived distance constraints, 104 dihedral angle restraints,26 distance restraints from hydrogen bonds.	XwinNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	300
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	sequential assignment bassed on triple resonance spectra, 2D 1H TOCSY, 3D 15N edited TOCSY, 3D 13C edited TOCSY

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	XwinNMR	2.6	Bruker
2	processing	XwinNMR	2.6	Bruker
3	data analysis	AUREMOL	2.2	Gronwald, Trenner, Brunner, Ganslmeier, Neidig & Kalbitzer
4	structure solution	CNS	1.1	Br nger
5	refinement	XPLOR-NIH	2.9.6	Schwieters
6	iterative matrix relaxation	AUREMOL	2.2	Gronwald, Trenner, Brunner, Ganslmeier, Neidig & Kalbitzer

RCSB PDB Core Operations are funded by the U.S. National Science Foundation (DBI-2321666), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM157729.