Experiment: 2KHX

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1-2 mM [U-99% 13C; U-99% 15N] protein, 25 mM [U-2H] TRIS, 100 mM potassium chloride, 1 mM [U-2H] DTT, 1 mM EDTA, 10 uM DSS	90% H2O/10% D2O	0.125	7.0	ambient	298
2	2D 1H-13C HSQC	1-2 mM [U-99% 13C; U-99% 15N] protein, 25 mM [U-2H] TRIS, 100 mM potassium chloride, 1 mM [U-2H] DTT, 1 mM EDTA, 10 uM DSS	90% H2O/10% D2O	0.125	7.0	ambient	298
3	3D HNCO	1-2 mM [U-99% 13C; U-99% 15N] protein, 25 mM [U-2H] TRIS, 100 mM potassium chloride, 1 mM [U-2H] DTT, 1 mM EDTA, 10 uM DSS	90% H2O/10% D2O	0.125	7.0	ambient	298
4	3D CBCA(CO)NH	1-2 mM [U-99% 13C; U-99% 15N] protein, 25 mM [U-2H] TRIS, 100 mM potassium chloride, 1 mM [U-2H] DTT, 1 mM EDTA, 10 uM DSS	90% H2O/10% D2O	0.125	7.0	ambient	298
5	3D HNCACB	1-2 mM [U-99% 13C; U-99% 15N] protein, 25 mM [U-2H] TRIS, 100 mM potassium chloride, 1 mM [U-2H] DTT, 1 mM EDTA, 10 uM DSS	90% H2O/10% D2O	0.125	7.0	ambient	298
6	3D H(CCO)NH	1-2 mM [U-99% 13C; U-99% 15N] protein, 25 mM [U-2H] TRIS, 100 mM potassium chloride, 1 mM [U-2H] DTT, 1 mM EDTA, 10 uM DSS	90% H2O/10% D2O	0.125	7.0	ambient	298
7	3D C(CO)NH	1-2 mM [U-99% 13C; U-99% 15N] protein, 25 mM [U-2H] TRIS, 100 mM potassium chloride, 1 mM [U-2H] DTT, 1 mM EDTA, 10 uM DSS	90% H2O/10% D2O	0.125	7.0	ambient	298
8	CN-4D simultaneous NOESY	1-2 mM [U-99% 13C; U-99% 15N] protein, 25 mM [U-2H] TRIS, 100 mM potassium chloride, 1 mM [U-2H] DTT, 1 mM EDTA, 10 uM DSS	90% H2O/10% D2O	0.125	7.0	ambient	298
9	HB-HE/HD-aromatic	1-2 mM [U-99% 13C; U-99% 15N] protein, 25 mM [U-2H] TRIS, 100 mM potassium chloride, 1 mM [U-2H] DTT, 1 mM EDTA, 10 uM DSS	90% H2O/10% D2O	0.125	7.0	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600

NMR Refinement
Method	Details	Software
simulated annealing	XPLOR-NIH, refine.py Loosened restraints after CYANA calculations	NMRPipe

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	chemical shift assignment	NMRView		Johnson, One Moon Scientific
3	structure solution	CYANA		Guntert, Mumenthaler and Wuthrich
4	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore

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.