2L9Q
Structural Characterization of small heat shock protein (Hsp12)
SOLUTION NMR
| NMR Experiment | ||||||||
|---|---|---|---|---|---|---|---|---|
| Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
| 1 | 2D 1H-15N HSQC | 0.2 mM [U-100% 15N] protein | 95% H2O/5% D2O | 100 | 7.0 | ambient | 313 | |
| 2 | 3D CBCA(CO)NH | 1.0 mM [U-100% 13C; U-100% 15N] protein | 95% H2O/5% D2O | 100 | 7.0 | ambient | 313 | |
| 3 | 3D HNCACB | 1.0 mM [U-100% 13C; U-100% 15N] protein | 95% H2O/5% D2O | 100 | 7.0 | ambient | 313 | |
| 4 | 2D 1H-13C HSQC aliphatic | 1.0 mM [U-100% 13C; U-100% 15N] protein | 95% H2O/5% D2O | 100 | 7.0 | ambient | 313 | |
| 5 | 2D 1H-13C HSQC aromatic | 1.0 mM [U-100% 13C; U-100% 15N] protein | 95% H2O/5% D2O | 100 | 7.0 | ambient | 313 | |
| 6 | 3D C(CO)NH | 1.0 mM [U-100% 13C; U-100% 15N] protein | 95% H2O/5% D2O | 100 | 7.0 | ambient | 313 | |
| 7 | 3D HBHA(CO)NH | 1.0 mM [U-100% 13C; U-100% 15N] protein | 95% H2O/5% D2O | 100 | 7.0 | ambient | 313 | |
| 8 | 3D HCCH-COSY | 1.0 mM [U-100% 13C; U-100% 15N] protein | 95% H2O/5% D2O | 100 | 7.0 | ambient | 313 | |
| 9 | 3D 1H-13C NOESY aliphatic | 1.0 mM [U-100% 13C; U-100% 15N] protein | 95% H2O/5% D2O | 100 | 7.0 | ambient | 313 | |
| 10 | 3D 1H-15N NOESY | 1.0 mM [U-100% 13C; U-100% 15N] protein | 95% H2O/5% D2O | 100 | 7.0 | ambient | 313 | |
| NMR Spectrometer Information | |||
|---|---|---|---|
| Spectrometer | Manufacturer | Model | Field Strength |
| 1 | Varian | INOVA | 900 |
| 2 | Varian | INOVA | 600 |
| NMR Refinement | ||
|---|---|---|
| Method | Details | Software |
| simulated annealing | CYANA | |
| NMR Ensemble Information | |
|---|---|
| Conformer Selection Criteria | target function |
| Conformers Calculated Total Number | 100 |
| Conformers Submitted Total Number | 20 |
| Representative Model | 1 (lowest energy) |
| Computation: NMR Software | ||||
|---|---|---|---|---|
| # | Classification | Version | Software Name | Author |
| 1 | chemical shift assignment | CYANA | Guntert, Mumenthaler and Wuthrich | |
| 2 | structure solution | MOLMOL | Koradi, Billeter and Wuthrich | |
| 3 | collection | VnmrJ | Varian | |
| 4 | processing | NMRPipe | Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax | |
| 5 | chemical shift assignment | PINE | Bahrami, Markley, Assadi, and Eghbalnia | |
| 6 | chemical shift assignment | TALOS | Cornilescu, Delaglio and Bax | |
| 7 | refinement | CYANA | Guntert, Mumenthaler and Wuthrich | |
