2N7L

NMR structure of the N-domain of troponin C bound to the switch region of troponin I and the covalent levosimendan analog i9

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.5-0.8 mM [U-15N] protein_1, 100 mM potassium chloride, 10 mM imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	95% H2O/5% D2O		6.9	ambient	303
2	2D 1H-13C HSQC	0.5-0.8 mM [U-13C; U-15N] protein_1, 100 mM potassium chloride, 10 mM imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	95% H2O/5% D2O		6.9	ambient	303
3	3D HNCACB	0.5-0.8 mM [U-13C; U-15N] protein_1, 100 mM potassium chloride, 10 mM imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	95% H2O/5% D2O		6.9	ambient	303
4	3D CBCA(CO)NH	0.5-0.8 mM [U-13C; U-15N] protein_1, 100 mM potassium chloride, 10 mM imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	95% H2O/5% D2O		6.9	ambient	303
5	3D HNHA	0.5-0.8 mM [U-15N] protein_1, 100 mM potassium chloride, 10 mM imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	95% H2O/5% D2O		6.9	ambient	303
6	3D H(CCO)NH	0.5-0.8 mM [U-13C; U-15N] protein_1, 100 mM potassium chloride, 10 mM imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	95% H2O/5% D2O		6.9	ambient	303
7	3D C(CO)NH	0.5-0.8 mM [U-13C; U-15N] protein_1, 100 mM potassium chloride, 10 mM imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	95% H2O/5% D2O		6.9	ambient	303
8	3D 1H-15N NOESY	0.5-0.8 mM [U-15N] protein_1, 100 mM potassium chloride, 10 mM imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	95% H2O/5% D2O		6.9	ambient	303
9	3D 1H-13C NOESY aliphatic	0.5-0.8 mM [U-13C; U-15N] protein_1, 100 mM potassium chloride, 10 mM [U-99% 2H] imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	100% D2O		6.9	ambient	303
10	gnoesyChsqc_CNfilt	0.5-0.8 mM [U-13C; U-15N] protein_1, 100 mM potassium chloride, 10 mM imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	95% H2O/5% D2O		6.9	ambient	303
11	2D 1H-1H NOESY CN filtered	0.5-0.8 mM [U-13C; U-15N] protein_1, 100 mM potassium chloride, 10 mM [U-99% 2H] imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	100% D2O		6.9	ambient	303
12	2D 1H-1H TOCSY	0.5-0.8 mM [U-13C; U-15N] protein_1, 100 mM potassium chloride, 10 mM [U-99% 2H] imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	100% D2O		6.9	ambient	303
13	2D 1H-19F HMQC	0.5-0.8 mM [U-13C; U-15N] protein_1, 100 mM potassium chloride, 10 mM [U-99% 2H] imidazole, 2 mM calcium chloride, 0.25 mM [U-99% 2H] DSS	100% D2O		6.9	ambient	303

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600
2	Varian	INOVA	800

NMR Refinement
Method	Details	Software
simulated annealing		VnmrJ

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	collection	VnmrJ		Varian
2	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	chemical shift assignment	NMRView		Johnson, One Moon Scientific
4	data analysis	NMRView		Johnson, One Moon Scientific
5	prediction of phi and psi dihedral angles	TALOS		Cornilescu, Delaglio and Bax
6	structure solution	X-PLOR NIH	2.35	Schwieters, Kuszewski, Tjandra and Clore
7	refinement	X-PLOR NIH	2.35	Schwieters, Kuszewski, Tjandra and Clore
8	generation of topology and parameter files for non-standard aminoacid ci9	PRODRG		A. W. Sch ttelkopf and D. M. F. van Aalten

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.