2KZ7
Solution structure of the CARMIL CAH3a/b domain bound to capping protein (CP)
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 | 0.35 mM [U-99% 15N; U-80% 2H] CPalpha subunit, 0.35 mM [U-99% 15N; U-80% 2H] CPbeta subunit, 0.39 mM [U-2H] CARMIL CAH3a/b domain | 93% H2O/7% D2O | 100 | 6.5 | ambient | 305 | |
2 | 2D 1H-15N HSQC | 0.35 mM [U-99% 15N; U-80% 2H] CARMIL CAH3a/b domain, 0.39 mM [U-2H] CPalpha subunit, 0.39 mM [U-2H] CPbeta subunit | 93% H2O/7% D2O | 100 | 6.5 | ambient | 305 | |
3 | 2D 1H-15N HSQC | 0.35 mM [U-99% 15N; U-80% 2H] CPalpha subunit, 0.35 mM [U-99% 15N; U-80% 2H] CPbeta subunit, 0.39 mM [U-2H; spin labeled at 1 or 5 postions] CARMIL CAH3a/b domain | 93% H2O/7% D2O | 100 | 6.5 | ambient | 305 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Bruker | DRX | 800 |
2 | Bruker | AVANCE | 900 |
NMR Refinement | ||
---|---|---|
Method | Details | Software |
molecular dynamics, simulated annealing, torsion angle dynamics | TopSpin |
NMR Ensemble Information | |
---|---|
Conformer Selection Criteria | structures with the lowest energy |
Conformers Calculated Total Number | 200 |
Conformers Submitted Total Number | 10 |
Representative Model | 1 (lowest energy) |
Computation: NMR Software | ||||
---|---|---|---|---|
# | Classification | Version | Software Name | Author |
1 | collection | TopSpin | Bruker Biospin | |
2 | processing | NMRPipe | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
3 | processing | NMRDraw | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
4 | chemical shift assignment | PIPP | Garrett | |
5 | chemical shift calculation | PIPP | Garrett | |
6 | peak picking | PIPP | Garrett | |
7 | structure solution | X-PLOR NIH | 2.23 | Schwieters, Kuszewski, Tjandra and Clore |
8 | refinement | X-PLOR NIH | 2.23 | Schwieters, Kuszewski, Tjandra and Clore |