SOLUTION NMR

NMR Experiment
Experiment Type Sample Contents Ionic Strength Solvent pH Pressure Temprature (K)
1 2D 1H-15N HSQC 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
2 2D 1H-15N HSQC 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
3 3D CBCA(CO)NH 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
4 3D CBCANH 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
5 3D HNCO 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
6 3D C(CO)NH 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
7 3D H(CCO)NH 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
8 3D HCCH-TOCSY 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
9 3D HCCH-COSY 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
10 3D HNHA 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
11 3D 1H-15N NOESY 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
12 3D 1H-13C NOESY 0.4-0.6 mM [U-98% 13C; U-98% 15N] copper binding protein, 10 % [U-99% 2H] D2O, 50 mM sodium phosphate, 50 mM sodium chloride, 5 mM EDTA, 5 mM TCEP, 10 uM DSS, 0.02 % sodium azide, 90% H2O/10% D2O 300 90% H2O/10% D2O 6 ambient 298
NMR Spectrometer Information
Spectrometer Manufacturer Model Field Strength
NMR Refinement
Method Details Software
simulated annealing, torsion angle dynamics ? 1
NMR Ensemble Information
Conformer Selection Criteria structures with the least restraint violations
Conformers Calculated Total Number 100
Conformers Submitted Total Number 18
Representative Model 1 (fewest violations)
Computation: NMR Software
# Classification Version Software Name Author
1 structure solution 2.1 CYANA Guntert, Mumenthaler and Wuthrich
2 peak picking 2.3.0 CcpNmr Analysis CCPN
3 data analysis 2.3.0 CcpNmr Analysis CCPN
4 chemical shift assignment 2.6 Sparky Goddard
5 peak picking 2.6 Sparky Goddard
6 processing 8.1 NMRPipe Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
7 structure validation 1.5 PSVS Bhattacharya and Montelione
8 chemical shift assignment 2.0 PINE Bahrami, Markley, Assadi, and Eghbalnia
9 refinement 2.36 X-PLOR NIH Schwieters, Kuszewski, Tjandra and Clore
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