Experiment: 7KYZ

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	1D 1H	0.7-0.9 mM [U-100% 15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III 700
2	2D 1H-15N HSQC	0.7-0.9 mM [U-100% 15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III 700
3	2D 1H-13C HSQC aliphatic	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III HD 800
4	2D 1H-13C HSQC aromatic	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III HD 800
5	3D 1H-15N TOCSY	0.7-0.9 mM [U-100% 15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III 700
15	3D HNCACB	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III 700
14	3D CBCA(CO)NH	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III 700
13	3D HNCA	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III 700
12	3D HN(CO)CA	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III 700
11	3D C(CO)NH	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III 700
10	3D HNCO	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE III 700
9	3D 1H-15N NOESY	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE 900
8	2D 1H-13C HSQC aromatic	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE 900
7	3D 1H-13C NOESY aliphatic	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE 900
6	2D 1H-15N HSQC	0.7-0.9 mM [U-13C; U-15N] GTPase KRas, 20 mM [U-2H] MES, 100 mM potassium chloride, 50 mM sodium chloride, 2 mM MgCl2, 1 mM [U-2H] TCEP, 7 mM [U-2H] D2O, 0.05 % sodium azide	90% H2O/10% D2O	0.15 M	6.5	1 atm	298	Bruker AVANCE 900

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

NMR Refinement
Method	Details	Software
torsion angle dynamics	structures are based on 2140 distance restraints, 138 hydrogen bond restraints as deduced from secondary structure and the tertiary fold detremined using refinement process, and 238 dihedral angle restraints	CYANA

NMR Ensemble Information
Conformer Selection Criteria	target function
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (target function)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin	4.0	Bruker Biospin
5	processing	NMRPipe	10.9	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	chemical shift assignment	CcpNmr Analysis	2.4	CCPN
3	peak picking	PONDEROSA		Lee W, Petit CM, Cornilescu G, Stark JL, Markley JL.
4	refinement	CYANA	3.98.13	Guntert, Mumenthaler and Wuthrich
6	refinement	PONDEROSA		Lee W, Petit CM, Cornilescu G, Stark JL, Markley JL.
7	structure calculation	CYANA	3.98.13	Guntert, Mumenthaler and Wuthrich

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 R01GM157729.