1Z60
Solution structure of the carboxy-terminal domain of human TFIIH P44 subunit
SOLUTION NMR
NMR Experiment | ||||||||
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Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
1 | 3D_15N-edited TOCSY and NOESY | 1mM p44 U-15N; 20mM deuterated TrisHCl buffer, 20mM NaCl and 0.5mM DTT | 90% H2O/10% D2O | 7 | ambient | 293 | ||
2 | 3D_15N-edited TOCSY and NOESY | 1mM p44 U-15N; 20mM deuterated TrisHCl buffer, 20mM NaCl and 0.5mM DTT | 90% H2O/10% D2O | 7 | ambient | 303 | ||
3 | 2D TOCSY and NOESY | 0.5mM p44; 20mM deuterated TrisHCl buffer, 20mM NaCl and 0.5mM DTT | 90% H2O/10% D2O | 7 | ambient | 303 | ||
4 | HSQC | 1mM p44 U-15N; 20mM deuterated TrisHCl buffer, 20mM NaCl and 0.5mM DTT | 90% H2O/10% D2O | 7 | ambient | 293 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Bruker | DRX | 600 |
NMR Refinement | ||
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Method | Details | Software |
simulated annealing | Automated assignment of the NOE was performed, The structures are based on 1294 unambiguous NOE-derived distance constraints, 32 dihedral angle restraints and 17 distance restraints from hydrogen bonds. Geometry of the zinc binding site II is distorded due to conformational exchange averaging. | ARIA / CNS |
NMR Ensemble Information | |
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Conformer Selection Criteria | structures with the lowest energy |
Conformers Calculated Total Number | 50 |
Conformers Submitted Total Number | 10 |
Representative Model | 1 (the best geometry) |
Additional NMR Experimental Information | |
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Details | 113Cd-1H HSQC spectrum was recorded using a broadband z-gradient probe at 293 K |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | structure solution | ARIA / CNS | 1.2. | Nilges |
2 | refinement | XPLOR-NIH | 2.0.6 | |
3 | data analysis | XEASY | 1.2 | Bartels |
4 | processing | XwinNMR | 2.6 | bruker |