1EGX | pdb_00001egx

SOLUTION STRUCTURE OF THE ENA-VASP HOMOLOGY 1 (EVH1) DOMAIN OF HUMAN VASODILATOR-STIMULATED PHOSPHOPROTEIN (VASP)

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_13C-separated_NOESY	1.3 mM VASP EVH1 DOMAIN 15N,13C; buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3; samples were used in both 90% H2O,10% D2O and 99.8% D2O 2 mM VASP EVH1 DOMAIN 15N buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3;sample were used in 90% H2O,10% D2O	90% H2O,10% D2O and 99.8% D2O	50 mM KCl, 20 mM KH2PO4	6.0	1 atm	300
2	3D_15N-separated_NOESY	1.3 mM VASP EVH1 DOMAIN 15N,13C; buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3; samples were used in both 90% H2O,10% D2O and 99.8% D2O 2 mM VASP EVH1 DOMAIN 15N buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3;sample were used in 90% H2O,10% D2O	90% H2O,10% D2O and 99.8% D2O	50 mM KCl, 20 mM KH2PO4	6.0	1 atm	300
3	2D NOESY	1.3 mM VASP EVH1 DOMAIN 15N,13C; buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3; samples were used in both 90% H2O,10% D2O and 99.8% D2O 2 mM VASP EVH1 DOMAIN 15N buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3;sample were used in 90% H2O,10% D2O	90% H2O,10% D2O and 99.8% D2O	50 mM KCl, 20 mM KH2PO4	6.0	1 atm	300
4	DQF-COSY	1.3 mM VASP EVH1 DOMAIN 15N,13C; buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3; samples were used in both 90% H2O,10% D2O and 99.8% D2O 2 mM VASP EVH1 DOMAIN 15N buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3;sample were used in 90% H2O,10% D2O	90% H2O,10% D2O and 99.8% D2O	50 mM KCl, 20 mM KH2PO4	6.0	1 atm	300
5	HNHA	1.3 mM VASP EVH1 DOMAIN 15N,13C; buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3; samples were used in both 90% H2O,10% D2O and 99.8% D2O 2 mM VASP EVH1 DOMAIN 15N buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3;sample were used in 90% H2O,10% D2O	90% H2O,10% D2O and 99.8% D2O	50 mM KCl, 20 mM KH2PO4	6.0	1 atm	300
6	3D HCCH-TOCSY	1.3 mM VASP EVH1 DOMAIN 15N,13C; buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3; samples were used in both 90% H2O,10% D2O and 99.8% D2O 2 mM VASP EVH1 DOMAIN 15N buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3;sample were used in 90% H2O,10% D2O	90% H2O,10% D2O and 99.8% D2O	50 mM KCl, 20 mM KH2PO4	6.0	1 atm	300
7	3D HNHB	1.3 mM VASP EVH1 DOMAIN 15N,13C; buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3; samples were used in both 90% H2O,10% D2O and 99.8% D2O 2 mM VASP EVH1 DOMAIN 15N buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3;sample were used in 90% H2O,10% D2O	90% H2O,10% D2O and 99.8% D2O	50 mM KCl, 20 mM KH2PO4	6.0	1 atm	300
8	3D_CBCA(CO)NNH	1.3 mM VASP EVH1 DOMAIN 15N,13C; buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3; samples were used in both 90% H2O,10% D2O and 99.8% D2O 2 mM VASP EVH1 DOMAIN 15N buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3;sample were used in 90% H2O,10% D2O	90% H2O,10% D2O and 99.8% D2O	50 mM KCl, 20 mM KH2PO4	6.0	1 atm	300
9	3D_CBCANNH	1.3 mM VASP EVH1 DOMAIN 15N,13C; buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3; samples were used in both 90% H2O,10% D2O and 99.8% D2O 2 mM VASP EVH1 DOMAIN 15N buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3;sample were used in 90% H2O,10% D2O	90% H2O,10% D2O and 99.8% D2O	50 mM KCl, 20 mM KH2PO4	6.0	1 atm	300
10	3D_15N-separated-TOCSY	1.3 mM VASP EVH1 DOMAIN 15N,13C; buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3; samples were used in both 90% H2O,10% D2O and 99.8% D2O 2 mM VASP EVH1 DOMAIN 15N buffer containing 20 mM KH2PO4, 50 mM KCl, 0.2 mM Na3N3;sample were used in 90% H2O,10% D2O	90% H2O,10% D2O and 99.8% D2O	50 mM KCl, 20 mM KH2PO4	6.0	1 atm	300

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	600
2	Bruker	DMX	750

NMR Refinement
Method	Details	Software
simulated annealing torsion angle dynamics	structures are based on 3043 NOE-derived distance restraints, 49 dihedral angle restraints, 25 distance restraints from hydrogen bonds	CNS

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations, structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	15
Representative Model	1 (fewest violations, lowest energy)

Additional NMR Experimental Information
Details	The structure was determined using triple-resonance NMR spectroscopy

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CNS	09a	Brunger
2	processing	Azara	2.1	Boucher
3	processing	XwinNMR	1.3	Bruker
4	collection	ANSIG	3.3	Kraulis
5	refinement	CNS	09a	Brunger

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.