Experiment: 2L4V

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
2	2D 1H-13C HSQC	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
3	3D HNCA	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
4	3D HN(CO)CA	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
5	3D HNCO	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
6	3D HN(CA)CO	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
7	3D CBCA(CO)NH	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
8	3D HNCACB	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
9	3D C(CO)NH	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
10	3D HBHA(CO)NH	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
11	3D H(CCO)NH	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
12	2D DQF-COSY	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
13	2D 1H-1H NOESY	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
14	3D HCCH-COSY	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
15	3D HCCH-TOCSY	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
16	3D 1H-15N TOCSY	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
17	3D 1H-15N NOESY	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303
18	3D 1H-13C NOESY	0.5 to 1 mM [U-99% 13C; U-99% 15N] Pineapple Cystatin-1	90% H2O/10% D2O	0.02	6.5	ambient	303

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

NMR Refinement
Method	Details	Software
simulated annealing	CNS	AURELIA

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	data analysis	AURELIA	3.9.7	Neidig, Geyer, Gorler, Antz, Saffrich, Beneicke, Kalbitzer
2	chemical shift assignment	AURELIA	3.9.7	Neidig, Geyer, Gorler, Antz, Saffrich, Beneicke, Kalbitzer
3	chemical shift assignment	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
4	calculation;	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
5	refinement	CNS	1.2	Brunger, Adams, Clore, Gros, Nilges and Read

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.