Experiment: 2L7A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1 mM [U-100% 15N] Vbs2b-1; 10 % [U-100% 2H] D2O-2; 50 mM sodium chloride-3; 2 mM DTT-4, 20 mM TRIS-5; 90% H2O/10% D2O	90% H2O/10% D2O	50	6.5	ambient	298
2	2D 1H-13C HSQC	1 mM [U-100% 13C; U-100% 15N] Vbs2b-6; 10 % [U-100% 2H] D2O-7; 50 mM sodium chloride-8; 2 mM DTT-9; 20 mM TRIS-10	90% H2O/10% D2O	50	6.5	ambient	298
3	2D 1H-1H NOESY	1 mM [U-100% 15N] Vbs2b-11, 100 % [U-100% 2H] D2O-12; 50 mM sodium chloride-13; 2 mM DTT-14; 20 mM TRIS-15; 100% D2O	100% D2O	50	6.5	ambient	298
4	3D CBCA(CO)NH	1 mM [U-100% 13C; U-100% 15N] Vbs2b-6; 10 % [U-100% 2H] D2O-7; 50 mM sodium chloride-8; 2 mM DTT-9; 20 mM TRIS-10	90% H2O/10% D2O	50	6.5	ambient	298
5	3D HCCH-TOCSY	1 mM [U-100% 13C; U-100% 15N] Vbs2b-6; 10 % [U-100% 2H] D2O-7; 50 mM sodium chloride-8; 2 mM DTT-9; 20 mM TRIS-10	90% H2O/10% D2O	50	6.5	ambient	298
6	3D HNCA	1 mM [U-100% 13C; U-100% 15N] Vbs2b-6; 10 % [U-100% 2H] D2O-7; 50 mM sodium chloride-8; 2 mM DTT-9; 20 mM TRIS-10	90% H2O/10% D2O	50	6.5	ambient	298
7	3D 1H-15N NOESY	1 mM [U-100% 15N] Vbs2b-1; 10 % [U-100% 2H] D2O-2; 50 mM sodium chloride-3; 2 mM DTT-4, 20 mM TRIS-5; 90% H2O/10% D2O	90% H2O/10% D2O	50	6.5	ambient	298
8	3D 1H-13C NOESY	1 mM [U-100% 13C; U-100% 15N] Vbs2b-6; 10 % [U-100% 2H] D2O-7; 50 mM sodium chloride-8; 2 mM DTT-9; 20 mM TRIS-10	90% H2O/10% D2O	50	6.5	ambient	298
9	3D HNCO	1 mM [U-100% 13C; U-100% 15N] Vbs2b-6; 10 % [U-100% 2H] D2O-7; 50 mM sodium chloride-8; 2 mM DTT-9; 20 mM TRIS-10	90% H2O/10% D2O	50	6.5	ambient	298

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

NMR Refinement
Method	Details	Software
simulated annealing, molecular dynamics, torsion angle dynamics	FINAL STRUCTURES REFINED IN EXPLICIT WATER BATH AS IMPLEMENTED IN ARIA 1.2/CNS 1.1. 20 LOWEST ENERGY STRUCTURES SELECTED FROM WATER REFINEMENT, CNS, INITIAL STRUCTURES GENERATED WITH CYANA	ARIA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	geometry optimization	ARIA	1.2	Linge, O'Donoghue and Nilges
2	structure solution	ARIA	1.2	Linge, O'Donoghue and Nilges
3	refinement	ARIA	1.2	Linge, O'Donoghue and Nilges
4	data analysis	CCPNAnalysis	1.15	CCPN
5	chemical shift assignment	CCPNAnalysis	1.15	CCPN
6	peak picking	CCPNAnalysis	1.15	CCPN
7	refinement	CNS	1.1	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.