1KLS
NMR Structure of the ZFY-6T[Y10L] Zinc Finger
SOLUTION NMR
NMR Experiment | ||||||||
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Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
1 | 2D NOESY | 2mM ZFY-6T[Y10L] | 90% H2O/10% D2O | 50mM d11-Tris-HCl, 2.2mM ZnCl2 | 6.0 | ambient | 298 | |
2 | 2D NOESY | 2mM ZFY-6T[Y10L] | 99.98% D2O. | 50mM d11-Tris-HCl, 2.2mM ZnCl2 | 6.0 | ambient | 298 | |
3 | DQF-COSY | 2mM ZFY-6T[Y10L] | 90% H2O/10% D2O | 50mM d11-Tris-HCl, 2.2mM ZnCl2 | 6.0 | ambient | 298 | |
4 | DQF-COSY | 2mM ZFY-6T[Y10L] | 99.98% D2O. | 50mM d11-Tris-HCl, 2.2mM ZnCl2 | 6.0 | ambient | 298 | |
5 | 3D-TOCSY-NOESY | 5mM ZFY-6T[Y10L] | 90% H2O/10% D2O | 50mM d11-Tris-HCl, 5.5mM ZnCl2 | 6.0 | ambient | 298 | |
6 | 2D-ROESY | 5mM ZFY-6T[Y10L] | 90% H2O/10% D2O | 50mM d11-Tris-HCl, 5.5mM ZnCl2 | 6.0 | ambient | 298 | |
7 | 2D-ROESY | 5mM ZFY-6T[Y10L] | 99.98% D2O. | 50mM d11-Tris-HCl, 5.5mM ZnCl2 | 6.0 | ambient | 298 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Varian | VXRS | 500 |
2 | Varian | UNITYPLUS | 500 |
3 | Varian | INOVA | 500 |
NMR Refinement | ||
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Method | Details | Software |
Distance geometry/Simulated annealing, followed by restrained molecular dynamics. The stereospecific assignments of the L10 methyl groups were determined by relaxation-matrix NOE backcalculation. | Structures are based on 265 interresidue NOE-derived distance constraints, 27 dihedral angle restraints, and 10 hydrogen bond restraints. | VNMR |
NMR Ensemble Information | |
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Conformer Selection Criteria | structures with the least restraint violations |
Conformers Calculated Total Number | 50 |
Conformers Submitted Total Number | 30 |
Representative Model | 6 (fewest violations) |
Additional NMR Experimental Information | |
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Details | This structure was determined using 2D and 3D homonuclear techniques.Stereospecific assignments for the L18 methyl groups could not be unambiguously determined. However, the resonances were clearly resolved. As a result,the structures were calculated as two families, depending on the assignments used. Structures 1-15 belong to one family, and use the assignments indicated in the accompanying restraint file. Structures 16-30 use the opposite assignments for the L18 methyls. |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | collection | VNMR | 4.3, 5.3 | Varian, Inc. |
2 | processing | VNMR | 4.3, 5.3 | Varian, Inc. |
3 | data analysis | NHFIT | Redfield, C. | |
4 | structure solution | DGII | standalone | Havel, T.F. |
5 | refinement | X-PLOR | 3.1 | Brunger, A.T. |