Experiment: 6YEL

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE III 950
2	2D 1H-13C HSQC	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE III 950
3	3D HNCO	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE III 950
4	3D HNCA	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE III 700
5	3D HNCACB	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE III 700
6	3D HN(CO)CA	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE III 700
8	3D 1H-15N TOCSY	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE III 950
7	3D 1H-15N NOESY	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE III 950
9	3D 1H-13C NOESY aliphatic	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE III 700
10	CON	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE NEO 700
11	CBCACON	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE NEO 700
12	CANCO	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE NEO 700
13	CACO	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE NEO 700
14	CBCACO	0.3 mM [U-98% 13C; U-98% 15N] STIM1 CC1, 20 mM TRIS, 7 mM [U-13C] SDS, 90 % v/v H2O, 10 % v/v [U-99% 2H] H2O	90% H2O/10% D2O	0.02 M	7.25	1 atm	310.15	Bruker AVANCE NEO 700

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE III	950
2	Bruker	AVANCE III	900
4	Bruker	AVANCE III	700
3	Bruker	AVANCE NEO	700

NMR Refinement
Method	Details	Software
simulated annealing		YASARA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	400
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin	3.5, 3.6, 4.0	Bruker Biospin
2	chemical shift assignment	CARA	1.8.4.2	Keller and Wuthrich
3	structure calculation	CS-ROSETTA		Shen, Vernon, Baker and Bax
4	structure calculation	CYANA		Guntert, Mumenthaler and Wuthrich
5	refinement	YASARA		Elmar Krieger Jacobus van Meel

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.