Experiment: 2N3T

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.7 mM [U-100% 13C; U-100% 15N; U-80% 2H] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide	95% H2O/5% D2O	50	6.7	ambient	298
2	2D 1H-13C HSQC	0.7 mM [U-100% 13C] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide	100% D2O	50	6.7	ambient	298
3	3D HNCACB	0.7 mM [U-100% 13C; U-100% 15N; U-80% 2H] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide	95% H2O/5% D2O	50	6.7	ambient	298
4	3D HNCA	0.7 mM [U-100% 13C; U-100% 15N; U-80% 2H] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide	95% H2O/5% D2O	50	6.7	ambient	298
5	3D HN(CO)CA	0.7 mM [U-100% 13C; U-100% 15N; U-80% 2H] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide	95% H2O/5% D2O	50	6.7	ambient	298
6	3D HNCO	0.7 mM [U-100% 13C; U-100% 15N; U-80% 2H] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide	95% H2O/5% D2O	50	6.7	ambient	298
7	3D HCCH-TOCSY	0.7 mM [U-100% 13C] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide	100% D2O	50	6.7	ambient	298
8	3D 1H-15N NOESY	0.7 mM [U-100% 15N] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide	95% H2O/5% D2O	50	6.7	ambient	298
9	3D 1H-13C NOESY aliphatic	0.7 mM [U-100% 13C] protein, 50 mM HEPES, 50 mM sodium chloride, 2 mM DTT, 1 mM EDTA, 0.1 % sodium azide	100% D2O	50	6.7	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	850
2	Bruker	AVANCE	900
3	Bruker	AVANCE	700
4	Varian	INOVA	800

NMR Refinement
Method	Details	Software
simulated annealing		TopSpin

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	50
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin		Bruker Biospin
2	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	chemical shift assignment	XEASY		Bartels et al.
4	peak picking	XEASY		Bartels et al.
5	data analysis	XEASY		Bartels et al.
6	data analysis	TALOS		Cornilescu, Delaglio and Bax
7	structure solution	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
8	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore

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 R01GM133198.