SOLUTION NMR

NMR Experiment
Experiment Type Sample Contents Ionic Strength Solvent pH Pressure Temprature (K)
1 1D 1H 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
2 2D 1H-15N HSQC 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
3 2D 1H-13C HSQC 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
4 3D HNCACB 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
5 3D CBCA(CO)NH 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
6 3D HNCA 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
7 3D HN(CO)CA 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
8 3D HNCO 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
9 3D HN(CA)CO 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
10 3D HBHA(CO)NH 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
11 3D H(CCO)NH 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
12 3D C(CO)NH 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
13 3D CCH-TOCSY 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
14 3D HCCH-TOCSY 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
15 3D HCCH-COSY 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
16 3D 15N-separated NOESY 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
17 3D 13C-separated NOESY 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
18 T1/R1 relaxation 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
19 T2/R2 relaxation 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
20 1H-15N heteronoe 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
21 2D 1H-15N HSQC NH2 only 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
22 T1rho/R1rho relaxation 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
23 2D HBCBCGCDCEHE 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
24 2D HBCBCGCDHD 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
25 3D 1H-13C NOESY aliphatic 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
26 3D 1H-13C NOESY aromatic 0.8 mM [U-100% 13C; U-100% 15N] KRAS4b protein, 0.02 % NaN3, 0.8 mM GppNHp, 1 mM TCEP, 100 mM KCl, 50 mM NaCl, 2 mM MgCl2, 20 mM MES, 250 uM DSS, 7 % [U-2H] D2O, 93 % H2O, 93% H2O/7% D2O 0.15 93% H2O/7% D2O 6.5 1 298
NMR Spectrometer Information
Spectrometer Manufacturer Model Field Strength
NMR Refinement
Method Details Software
torsion angle dynamics ? 6
NMR Ensemble Information
Conformer Selection Criteria target function
Conformers Calculated Total Number 100
Conformers Submitted Total Number 20
Representative Model 1 (lowest energy & target function)
Computation: NMR Software
# Classification Version Software Name Author
1 processing v10.9 NMRPipe Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2 data analysis v10.9 NMRDraw Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3 chemical shift assignment v2.4 CCNMR Vranken WF, Boucher W, Stevens TJ, Fogh RH, Pajon A, Llinas M, Ulrich EL, Markley JL, Ionides J, and Laue ED
5 structure calculation ? CYANA Guntert, Mumenthaler and Wuthrich
6 refinement 1.3 CNS Brunger, Adams, Clore, Gros, Nilges and Read
7 peak picking ? CcpNmr Analysis CCPN
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