2IFS

Structure of the N-WASP EVH1 domain in complex with an extended WIP peptide

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_15N-separated_NOESY	1.0 mM U-15N/13C protein, 20 mM sodium phosphate, 1 mM Dithiothreitol, 90% H2O, 10% D2O	90% H2O/10% D2O	0.044 mM	7.0	1 atm	303
2	3D_13C-separated_NOESY	1.0 mM U-15N/13C protein, 20 mM sodium phosphate, 1 mM Dithiothreitol, 90% H2O, 10% D2O	90% H2O/10% D2O	0.044 mM	7.0	1 atm	303
3	3D_13C-separated_NOESY (AROMATIC)	1.0 mM U-15N/13C protein, 20 mM sodium phosphate, 1 mM Dithiothreitol, 90% H2O, 10% D2O	90% H2O/10% D2O	0.044 mM	7.0	1 atm	303

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	600

NMR Refinement
Method	Details	Software
AUTOMATED METHODS WERE USED FOR BACKBONE CHEMICAL SHIFT ASSIGNMENT AND ITERATIVE NOE REFINEMENT. FINAL STRUCTURES WERE OBTAINED BY MOLECULAR DYNAMICS IN EXPLICIT SOLVENT	STRUCTURES ARE BASED ON A TOTAL OF 2411 NOE CONSTRAINTS ( 383 INTRA, 473 SEQUENTIAL, 386 MEDIUM and 922 INTRAMOLECULAR LONG RANGE - EVH1 DOMAIN, 247 INTERMOLECULAR WIP-EVH1 CONSTRAINTS) AND 253 PHI AND PSI DIHEDRAL ANGLE CONSTRAINTS.	Xplor-NIH

NMR Ensemble Information
Conformer Selection Criteria	target function
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	Xplor-NIH	2.9.3	SCHWIETERS,C.D.,KUSZEWSKI,J.J.,TJANDRA,N.,CLORE,G.M.
2	collection	XwinNMR	3.5	Bruker
3	processing	NMRPipe	2004	Delagio,F. et al.
4	data analysis	XEASY	1.3	Eccles, C., Guntert, P., Billeter, M., Wuthrich, K.
5	data analysis	SPSCAN	1.1.0	R.W. Glaser
6	data analysis	GARANT	2.1	C. Bartels
7	structural calculation	CYANA	2.1	Guntert, P.

RCSB PDB Core Operations are funded by the National Science Foundation (DBI-1832184), 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 R01GM133198.