Experiment: 2K60

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.5-1.0 mM [U-100% 13C; U-100% 15N] stromal interaction molecule 1, 20 mM TRIS, 100 mM sodium chloride, 5 mM CALCIUM ION	95% H2O/5% D2O	105	8.0	1 atm	293
2	3D CBCA(CO)NH	0.5-1.0 mM [U-100% 13C; U-100% 15N] stromal interaction molecule 1, 20 mM TRIS, 100 mM sodium chloride, 5 mM CALCIUM ION	95% H2O/5% D2O	105	8.0	1 atm	293
3	3D HNCACB	0.5-1.0 mM [U-100% 13C; U-100% 15N] stromal interaction molecule 1, 20 mM TRIS, 100 mM sodium chloride, 5 mM CALCIUM ION	95% H2O/5% D2O	105	8.0	1 atm	293
4	3D HNCO	0.5-1.0 mM [U-100% 13C; U-100% 15N] stromal interaction molecule 1, 20 mM TRIS, 100 mM sodium chloride, 5 mM CALCIUM ION	95% H2O/5% D2O	105	8.0	1 atm	293
5	3D H(CCO)NH	0.5-1.0 mM [U-100% 13C; U-100% 15N] stromal interaction molecule 1, 20 mM TRIS, 100 mM sodium chloride, 5 mM CALCIUM ION	95% H2O/5% D2O	105	8.0	1 atm	293
6	3D C(CO)NH	0.5-1.0 mM [U-100% 13C; U-100% 15N] stromal interaction molecule 1, 20 mM TRIS, 100 mM sodium chloride, 5 mM CALCIUM ION	95% H2O/5% D2O	105	8.0	1 atm	293
7	3D 1H-15N NOESY	0.5-1.0 mM [U-100% 13C; U-100% 15N] stromal interaction molecule 1, 20 mM TRIS, 100 mM sodium chloride, 5 mM CALCIUM ION	95% H2O/5% D2O	105	8.0	1 atm	293
8	3D HCCH-TOCSY	0.5-1.0 mM [U-100% 13C; U-100% 15N] stromal interaction molecule 1, 20 mM [U-99% 2H] TRIS, 100 mM sodium chloride, 5 mM CALCIUM ION	100% D2O	105		1 atm	293
9	2D 1H-13C HSQC	0.5-1.0 mM [U-100% 13C; U-100% 15N] stromal interaction molecule 1, 20 mM [U-99% 2H] TRIS, 100 mM sodium chloride, 5 mM CALCIUM ION	100% D2O	105		1 atm	293
10	3D 1H-13C NOESY	0.5-1.0 mM [U-100% 13C; U-100% 15N] stromal interaction molecule 1, 20 mM [U-99% 2H] TRIS, 100 mM sodium chloride, 5 mM CALCIUM ION	100% D2O	105		1 atm	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	500
2	Bruker	Avance	800

NMR Refinement
Method	Details	Software
simulated annealing, simulated annealing	Water refinement and violation analysis was performed using the RECOORD scripts in CNS v1.1 (Nederveen et al, 2005).	CNS

NMR Ensemble Information
Conformer Selection Criteria	20 structures for lowest energy
Conformers Calculated Total Number	250
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	CNS	v1.1	Brunger A. T. et.al.
2	structure solution	CYANA	v2.1	Guntert, Mumenthaler and Wuthrich
3	chemical shift assignment	XEASY		Bartels et al.
4	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax

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.