Experiment: 6VK2

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III HD 900
2	3D 1H-13C NOESY aliphatic	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III HD 900
3	3D 1H-13C NOESY aromatic	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III HD 900
4	3D 1H-15N NOESY	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III HD 900
5	3D HNCACB	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III 800
6	3D HNCA	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III HD 700
7	3D CBCA(CO)NH	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III 800
8	3D HBHA(CO)NH	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III 800
9	3D HNCO	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III 800
10	3D H(CCO)NH	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III 800
11	3D HCCH-TOCSY	200 mM sodium chloride, 20 mM sodium phosphate, 1 mM EDTA, 1 mM PMSF, 5 mM sodium azide, 300 uM [U-100% 13C; U-100% 15N] Grb2-SH2	90% H2O/10% D2O	138.2 mM	7	1 atm	298	Bruker AVANCE III 800

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE III HD	900
2	Bruker	AVANCE III	800
3	Bruker	AVANCE III HD	700

NMR Refinement
Method	Details	Software
na		CS-ROSETTA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	66317
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure calculation	CS-ROSETTA	3.3	Shen, Vernon, Baker and Bax
2	data analysis	CcpNmr Analysis		CCPN
3	structure calculation	ARIA		Linge, O'Donoghue and Nilges
4	refinement	CS-ROSETTA		Shen, Vernon, Baker and Bax
5	chemical shift assignment	CcpNmr Analysis		CCPN
6	peak picking	CcpNmr Analysis		CCPN

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.