Experiment: 5UKE

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
2	2D 1H-13C HSQC	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
3	3D CBCA(CO)NH	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
4	3D C(CO)NH	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
5	3D HNCO	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
6	3D HNCA	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
7	3D HNCACB	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
8	3D HN(CO)CA	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
12	3D H(CCO)NH	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
11	3D HCCH-TOCSY	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
10	3D 1H-15N NOESY	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
9	3D 1H-15N TOCSY	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
13	3D 1H-13C NOESY aliphatic	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600
14	3D 1H-13C NOESY aromatic	800 uM [U-99% 13C; U-99% 15N] IRAK-M Death Domain_R56D_Y61E, 20 mM sodium chloride, 5 mM TCEP, 10 mM TRIS, 5 mM sodium azide	90% H2O/10% D2O	0.0229 M	6.67	1 atm	298	Varian INOVA 600

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

NMR Refinement
Method	Details	Software
DGSA-distance geometry simulated annealing	NIH-Xplor imbedded to Ponderosa	PONDEROSA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	chemical shift assignment	PINE	2.0	Bahrami, Markley, Assadi, and Eghbalnia
2	data analysis	Sparky	3.115	Goddard
3	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	refinement	PONDEROSA		NMRFAM (University of Wisconsin-Madison), WoongHee Lee
5	structure calculation	PONDEROSA		NMRFAM (University of Wisconsin-Madison), WoongHee Lee

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.