Experiment: 5K6P

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
2	3D HNCACB	412 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
4	3D HNCO	412 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 600
1	2D 1H-15N HSQC	920 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
16	2D 1H-13C HSQC	920 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
3	HCC(CO)NH TOCSY	920 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
8	CC(CO)NH-TOCSY	920 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
7	3D HBHA(CO)NH	920 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
9	3D CBCA(CO)NH	920 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
12	3D 1H-15N NOESY	550 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 1 mM Na azide, 2 mM TCEP, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
10	3D 13C NOESY aromatic	550 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 1 mM Na azide, 2 mM TCEP, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
6	3D HCCH-TOCSY	550 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 1 mM Na azide, 2 mM TCEP, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
20	3D HCCH-TOCSY	550 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 1 mM Na azide, 2 mM TCEP, 10 mM DSS	95% H2O/5% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
15	2D HBCBCGCDHD	500 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	99% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
5	2D HBCBCGCDCEHE	500 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	99% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 600
13	3D 13C NOESY aliphatic	500 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	99% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
11	2D 1H-13C HSQC aromatic	800 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	99% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
14	3D HCC TOCSY aromatic	800 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	99% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800
19	3D HCCH-TOCSY	800 uM [U-99% 13C; U-99% 15N] human cMyBP-C (319-451), 10 mM MES, 1 mM CaCl2, 50 mM NaCl, 2 mM TCEP, 1 mM Na azide, 10 mM DSS	99% D2O	50 mM	6.5	1 atm	298	Bruker AVANCE 800

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	600
2	Bruker	AVANCE	800

NMR Refinement
Method	Details	Software
molecular dynamics	torsion angle dynamics, water refinement was carried out using the lowest 50 energy structures from CYANA	CYANA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	500
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
2	refinement	CYANA		Guntert, Mumenthaler and Wuthrich
5	refinement	CNS	1.2	Brunger, Adams, Clore, Gros, Nilges and Read
3	chemical shift assignment	Analysis		CCPN
4	peak picking	Analysis		CCPN
6	structure calculation	CYANA		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.