2KJ6
NMR Solution Structure of a Tubulin folding cofactor B obtained from Arabidopsis thaliana: Northeast Structural Genomics Consortium target AR3436A
SOLUTION NMR
NMR Experiment | ||||||||
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Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
1 | 3D 1H-15N NOESY | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
2 | 3D 1H-13Caliphatic NOESY | 0.78 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
3 | 3D 1H-13C aromatic NOESY | 0.78 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
4 | 2D 1H-15N HSQC | 0.78 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
5 | 2D 1H-13C HSQC | 0.78 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
6 | 2D 1H-15N HSQC | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
7 | 2D 1H-13C HSQC | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
8 | 3D HBHA(CO)NH | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
9 | 3D CBCA(CO)NH | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
10 | (4,3)D GFT-HNNCABCA | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
11 | (4,3)D GFT-CABCA(CO)NHN | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
12 | 3D HNCO | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
13 | 3D HNCA | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
14 | 3D HNCACB | 0.95 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
15 | 3D HNHA | 0.56 mM [U-10% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
16 | 3D H(CCO)NH | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 | |
17 | 3D HCCH-TOCSY | 0.4 mM [U-100% 13C; U-100% 15N] AR3436A | 90% H2O/10% D2O | 5mM CaCl2, 10mM NaCl | 6.5 | ambient | 293 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Bruker | AVANCE | 800 |
2 | Varian | INOVA | 600 |
3 | Varian | INOVA | 500 |
NMR Refinement | ||
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Method | Details | Software |
simulated annealing | The structure was obtained using triple resonance NMR spectroscopy. GFT-NMR experiments were used for backbone assignments. Automated NOESY assignments were made using AUTOSTRUCTURE and CYANA-2.1. Dihedral angle constraints were obtained from TALOS. The assignments were validated using AVS software. Final structure factors determined using PSVS: ordered residues: 29-33, 15-21, 89-94, 57-62, 39-50, 79-82. Ramachandran statistics for ordered residues: Most favored region: 84.4%, additionally favored region: 15.6%, generously aloowed: 0.1%. Procheck scores for ordered residues (RAW/Z): Phi/psi -0.51/-1.69, all -0.33/-1.95, MolProbity clash scores (RAW/Z) - 15.41/-1.12. RPF scores for the goodness fit to NOESY data: Recall-0.993, Precision-0.878, F-measure-0.932, final dp score-0.756. | CNS |
NMR Ensemble Information | |
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Conformer Selection Criteria | structures with the lowest energy |
Conformers Calculated Total Number | 140 |
Conformers Submitted Total Number | 20 |
Representative Model | 1 (lowest energy) |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | refinement | CNS | 2.0.6 | Brunger, Adams, Clore, Gros, Nilges and Read |
2 | structure solution | AutoStructure | 2.2.1 | Huang, Tejero, Powers and Montelione |
3 | structure solution | CYANA | 2.1 | Guntert, Mumenthaler and Wuthrich |
4 | chemical shift assignment | AutoAssign | 2.4.0 | Zimmerman, Moseley, Kulikowski and Montelione |