6DMP
De Novo Design of a Protein Heterodimer with Specificity Mediated by Hydrogen Bond Networks
SOLUTION NMR
NMR Experiment | ||||||||
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Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
1 | 2D 1H-15N HSQC | 1.0 mM [U-13C; U-15N] DHD13_XAAA, 1.0 mM [U-13C; U-15N] DHD13_XAAB. 90% H2O/10% D2O | 90% H2O/10% D2O | 0.1 M | 7.2 | 1 atm | 308 | Bruker AVANCE III 800 |
2 | 2D 1H-13C HSQC | 1.0 mM [U-13C; U-15N] DHD13_XAAA, 1.0 mM [U-13C; U-15N] DHD13_XAAB. 90% H2O/10% D2O | 90% H2O/10% D2O | 0.1 M | 7.2 | 1 atm | 308 | Bruker AVANCE III 800 |
3 | 4D HNCH TOCSY | 1.0 mM [U-13C; U-15N] DHD13_XAAA, 1.0 mM [U-13C; U-15N] DHD13_XAAB. 90% H2O/10% D2O | 90% H2O/10% D2O | 0.1 M | 7.2 | 1 atm | 308 | Bruker AVANCE III 800 |
4 | 4D HNCH NOESY | 1.0 mM [U-13C; U-15N] DHD13_XAAA, 1.0 mM [U-13C; U-15N] DHD13_XAAB. 90% H2O/10% D2O | 90% H2O/10% D2O | 0.1 M | 7.2 | 1 atm | 308 | Bruker AVANCE III 800 |
5 | 4D HCCH NOESY | 1.0 mM [U-13C; U-15N] DHD13_XAAA, 1.0 mM [U-13C; U-15N] DHD13_XAAB. 90% H2O/10% D2O | 90% H2O/10% D2O | 0.1 M | 7.2 | 1 atm | 308 | Bruker AVANCE III 800 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Bruker | AVANCE III | 800 |
NMR Refinement | ||
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Method | Details | Software |
simulated annealing | CS-ROSETTA |
NMR Ensemble Information | |
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Conformer Selection Criteria | structures with the least restraint violations |
Conformers Calculated Total Number | 10000 |
Conformers Submitted Total Number | 10 |
Representative Model | 1 (lowest energy) |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | refinement | CS-ROSETTA | 3.0 | Shen, Vernon, Baker and Bax |
2 | chemical shift assignment | Sparky | Goddard | |
3 | peak picking | Sparky | Goddard | |
4 | processing | NMRPipe | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
5 | structure calculation | CS-ROSETTA | 3.0 | Shen, Vernon, Baker and Bax |