1T84

Solution structure of the Wiskott-Aldrich Syndrome Protein (WASP) autoinhibited core domain complexed with (S)-wiskostatin, a small molecule inhibitor

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	HCC-TOCSY	1mM WASP U-15N,13C; 1mM (S)-wiskostatin	20 mM Acetate-d3 pH 5.0; 20mM NaCl; 1mM DTT-d10; 5% DMSO-d6; 90% H2O/10% D2O or 100% D2O	20mM	5.0	ambient	298
2	CCC-TOCSY	1mM WASP U-15N,13C; 1mM (S)-wiskostatin	20 mM Acetate-d3 pH 5.0; 20mM NaCl; 1mM DTT-d10; 5% DMSO-d6; 90% H2O/10% D2O or 100% D2O	20mM	5.0	ambient	298
3	HCCH-TOCSY	1mM WASP U-15N,13C; 1mM (S)-wiskostatin	20 mM Acetate-d3 pH 5.0; 20mM NaCl; 1mM DTT-d10; 5% DMSO-d6; 90% H2O/10% D2O or 100% D2O	20mM	5.0	ambient	298
4	3D-13C-separated_NOESY_aliphatic	1mM WASP U-15N,13C; 1mM (S)-wiskostatin	20 mM Acetate-d3 pH 5.0; 20mM NaCl; 1mM DTT-d10; 5% DMSO-d6; 90% H2O/10% D2O or 100% D2O	20mM	5.0	ambient	298
5	3D-13C-separated_NOESY_aromatic	1mM WASP U-15N,13C; 1mM (S)-wiskostatin	20 mM Acetate-d3 pH 5.0; 20mM NaCl; 1mM DTT-d10; 5% DMSO-d6; 90% H2O/10% D2O or 100% D2O	20mM	5.0	ambient	298
6	3D_15N-separated_NOESY	1mM WASP U-15N,13C; 1mM (S)-wiskostatin	20 mM Acetate-d3 pH 5.0; 20mM NaCl; 1mM DTT-d10; 5% DMSO-d6; 90% H2O/10% D2O or 100% D2O	20mM	5.0	ambient	298
7	4D_13C-separated_NOESY	1mM WASP U-15N,13C; 1mM (S)-wiskostatin	20 mM Acetate-d3 pH 5.0; 20mM NaCl; 1mM DTT-d10; 5% DMSO-d6; 90% H2O/10% D2O or 100% D2O	20mM	5.0	ambient	298
8	4D_13C/15N-separated_NOESY	1mM WASP U-15N,13C; 1mM (S)-wiskostatin	20 mM Acetate-d3 pH 5.0; 20mM NaCl; 1mM DTT-d10; 5% DMSO-d6; 90% H2O/10% D2O or 100% D2O	20mM	5.0	ambient	298
9	3D_13C-edited_12C-filtered_NOESY	1mM WASP U-15N,13C; 1mM (S)-wiskostatin	20 mM Acetate-d3 pH 5.0; 20mM NaCl; 1mM DTT-d10; 5% DMSO-d6; 90% H2O/10% D2O or 100% D2O	20mM	5.0	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600

NMR Refinement
Method	Details	Software
simulated annealing; torsion angle dynamics	The structures are based on a total of 1466 restraints. 1298 are NOE-derived intramolecular distance restraints; 30 are NOE-derived intermolecular distance restraints; 138 are dihedral angle restraints.	NMRPipe

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

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	NMRPipe	1.7	Bax
2	data analysis	NMRView	2.1.2	Johnson
3	data analysis	VNMR	6.1B
4	structure solution	X-PLOR	3.851	Brunger
5	structure solution	ARIA	1	Nilges
6	refinement	ARIA	1	Nilges

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.