1SBJ

NMR Structure of the Mg2+-loaded C Terminal Domain of Cardiac Troponin C Bound to the N Terminal Domain of Cardiac Troponin I

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	HSQC	1mM [13C,15]cTnC(81-161)/cTnI(33-80), 20mM Tris(d11), 10mM DTT, 20mM beta-mercaptoethanol, 2mM EGTA, 100mM KCl, 20mM MgCl2	90% H20, 10% D2O	100 mM CaCl2, 20 mM MgCl2	6.8	ambient	318.00
2	HNHA, HNHB, CN-NOESY-HSQC	1mM [13C,15]cTnC(81-161)/cTnI(33-80), 20mM Tris(d11), 10mM DTT, 20mM beta-mercaptoethanol, 2mM EGTA, 100mM KCl, 20mM MgCl2	90% H20, 10% D2O	100 mM CaCl2, 20 mM MgCl2	6.8	ambient	318.00
3	(HB)CBCA(CO)NH, HNCACB	1mM [13C,15]cTnC(81-161)/cTnI(33-80), 20mM Tris(d11), 10mM DTT, 20mM beta-mercaptoethanol, 2mM EGTA, 100mM KCl, 20mM MgCl2	90% H20, 10% D2O	100 mM CaCl2, 20 mM MgCl2	6.8	ambient	318.00
4	H(CCO)NH,C(CO)NH	1mM [13C,15]cTnC(81-161)/cTnI(33-80), 20mM Tris(d11), 10mM DTT, 20mM beta-mercaptoethanol, 2mM EGTA, 100mM KCl, 20mM MgCl2	90% H20, 10% D2O	100 mM CaCl2, 20 mM MgCl2	6.8	ambient	318.00
5	(HB)CB(CGCD)HD, (HB)CB(CGCDCE)HE	1mM [13C,15]cTnC(81-161)/cTnI(33-80), 20mM Tris(d11), 10mM DTT, 20mM beta-mercaptoethanol, 2mM EGTA, 100mM KCl, 20mM MgCl2	90% H20, 10% D2O	100 mM CaCl2, 20 mM MgCl2	6.8	ambient	318.00
6	NOESY-HSQC, AB-TROSY	1mM [13C,15]cTnC(81-161)/cTnI(33-80), 20mM Tris(d11), 10mM DTT, 20mM beta-mercaptoethanol, 2mM EGTA, 100mM KCl, 20mM MgCl2	90% H20, 10% D2O	100 mM CaCl2, 20 mM MgCl2	6.8	ambient	318.00

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

NMR Refinement
Method	Details	Software
DGSA	the structures are based on a total of 958 restraints, 818 are NOE-derived distance restraints, 78 dihedral angle restraints, 4 distance restraints from hydrogen bonds and 58 dipolar coupling restraints	Felix

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	20
Conformers Submitted Total Number	20
Representative Model	11 (lowest energy)

Additional NMR Experimental Information
Details	Best 20 Structures. These structures were determined using triple-resonance NMR spectroscopy on Mg2+-saturated 15N,13C-cTnC(81-161) bound to cTnI(33-80).

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	Felix	2000	Accelrys
2	structure solution	CNS	1.0	Brunger
3	structure solution	CNX	1.0	Accelrys
4	refinement	CNX	1.0	Accelrys

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.