Experiment: 2NCP

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	90% H2O/10% D2O	0.05	7.5	ambient	298
2	3D HNCACB	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	90% H2O/10% D2O	0.05	7.5	ambient	298
3	3D CBCA(CO)NH	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	90% H2O/10% D2O	0.05	7.5	ambient	298
4	3D HNCO	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	90% H2O/10% D2O	0.05	7.5	ambient	298
5	3D H(CCO)NH	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	90% H2O/10% D2O	0.05	7.5	ambient	298
6	3D HCACO	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	100% D2O	0.05	7.5	ambient	298
7	2D 1H-13C HMQC	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	100% D2O	0.05	7.5	ambient	298
8	3D HCCH-TOCSY	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	100% D2O	0.05	7.5	ambient	298
9	3D 1H-13C NOESY	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	100% D2O	0.05	7.5	ambient	298
10	3D 1H-15N NOESY	0.75 mM [U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	90% H2O/10% D2O	0.05	7.5	ambient	298
11	2D 1H-1H COSY	0.75 mM Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	100% D2O	0.05	7.5	ambient	298
12	2D 1H-1H TOCSY	0.75 mM Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	100% D2O	0.05	7.5	ambient	298
13	2D 1H-1H NOESY	0.75 mM Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	100% D2O	0.05	7.5	ambient	298
14	2D (HB)CB(CGCD)HD	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	100% D2O	0.05	7.5	ambient	298
15	2D (HB)CB(CGCDCE)HE	0.75 mM [U-98% 13C; U-98% 15N] Tcb2-C, 25 mM TRIS, 50 mM potassium chloride, 5 mM magnesium chloride, 1 mM EGTA, 5 mM calcium chloride	100% D2O	0.05	7.5	ambient	298

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

NMR Refinement
Method	Details	Software
simulated annealing		NMRPipe

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	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	data analysis	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	collection	VnmrJ		Varian
4	peak picking	Analysis		CCPN
5	data analysis	Analysis		CCPN
6	chemical shift assignment	Analysis		CCPN
7	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
8	refinement	CYANA	2.1	Guntert, Mumenthaler and Wuthrich

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.