Experiment: 2KMC

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.4mM F0-1, 10% [U-100% 2H] D2O-2, 2mM DTT-3, 50mM sodium chloride-4, 20mM sodium phosphate-5	90% H2O/10% D2O	50	6.5	ambient	298
2	2D 1H-13C HSQC	0.4mM [U-100% 13C; U-100% 15N] F0-6, 10% [U-100% 2H] D2O-7, 2mM DTT-8, 50 mM sodium chloride-9, 20mM sodium phosphate-10	90% H2O/10% D2O	50	6.5	ambient	298
3	3D CBCA(CO)NH	0.4mM [U-100% 13C; U-100% 15N] F0-6, 10% [U-100% 2H] D2O-7, 2mM DTT-8, 50 mM sodium chloride-9, 20mM sodium phosphate-10	90% H2O/10% D2O	50	6.5	ambient	298
4	3D HNCO	0.4mM [U-100% 13C; U-100% 15N] F0-6, 10% [U-100% 2H] D2O-7, 2mM DTT-8, 50 mM sodium chloride-9, 20mM sodium phosphate-10	90% H2O/10% D2O	50	6.5	ambient	298
5	3D HNCA	0.4mM [U-100% 13C; U-100% 15N] F0-6, 10% [U-100% 2H] D2O-7, 2mM DTT-8, 50 mM sodium chloride-9, 20mM sodium phosphate-10	90% H2O/10% D2O	50	6.5	ambient	298
6	3D HNCACB	0.4mM [U-100% 13C; U-100% 15N] F0-6, 10% [U-100% 2H] D2O-7, 2mM DTT-8, 50 mM sodium chloride-9, 20mM sodium phosphate-10	90% H2O/10% D2O	50	6.5	ambient	298
7	3D HCCH-TOCSY	0.4mM [U-100% 13C; U-100% 15N] F0-6, 10% [U-100% 2H] D2O-7, 2mM DTT-8, 50 mM sodium chloride-9, 20mM sodium phosphate-10	90% H2O/10% D2O	50	6.5	ambient	298
8	3D 1H-15N NOESY	0.4mM F0-1, 10% [U-100% 2H] D2O-2, 2mM DTT-3, 50mM sodium chloride-4, 20mM sodium phosphate-5	90% H2O/10% D2O	50	6.5	ambient	298
9	3D 1H-13C NOESY	0.4mM [U-100% 13C; U-100% 15N] F0-6, 10% [U-100% 2H] D2O-7, 2mM DTT-8, 50 mM sodium chloride-9, 20mM sodium phosphate-10	90% H2O/10% D2O	50	6.5	ambient	298
10	2D 1H-1H NOESY	0.4mM F0-11, 100% [U-100% 2H] D2O-12, 2mM DTT-13, 50mM sodium chloride-14, 20mM sodium phosphate-15	100% D2O	50	6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	600
2	Bruker	DRX	600
3	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 using CNS. Initial Structures generated with cyana	TopSpin

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	collection	TopSpin	2.0	Bruker Biospin
2	processing	TopSpin	2.0	Bruker Biospin
3	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
4	chemical shift assignment	Analysis	1.0.15	CCPN
5	data analysis	Analysis	1.0.15	CCPN
6	peak picking	Analysis	1.0.15	CCPN
7	refinement	CNS	1.1	Brunger, Adams, Clore, Gros, Nilges and Read
8	refinement	ARIA	1.2	Linge, O'Donoghue and Nilges
9	refinement	CYANA	2.1	Guntert, Mumenthaler and Wuthrich

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.