2L1R

The structure of the calcium-sensitizer, dfbp-o, in complex with the N-domain of troponin C and the switch region of troponin I

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
2	2D 1H-13C HSQC	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
3	3D CBCA(CO)NH	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
4	3D C(CO)NH	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
5	3D H(CCO)NH	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
6	3D HNCACB	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
7	3D 1H-13C NOESY	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
8	3D 1H-15N NOESY	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
9	3D 1H-15N TOCSY	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
10	2D 13C, 15N Filtered NOESY	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
11	2D 13C,15N Filtered TOCSY	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
12	2D 1H-19F HOESY	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303
13	3D 13C filtered, edited NOESY	0.5-1 mM [U-99% 13C; U-99% 15N] cNTnC, 2-4 mM cTnI(144-163), 2-4 mM 2',4'-difluoro (1,1'-biphenyl)-4-yloxy acetic acid, 2-4 mM Calcium, 100 mM KCl, 10 mM DTT, 10 mM Imidazole	95% H2O/5% D2O	0.1	6.8	ambient	303

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	500
2	Varian	UNITY	600

NMR Refinement
Method	Details	Software
simulated annealing		X-PLOR NIH

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	200
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	X-PLOR NIH	2.24	Schwieters, Kuszewski, Tjandra and Clore

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.