2OSG
Solution Structure and Binding Property of the Domain-swapped Dimer of ZO2PDZ2
SOLUTION NMR
NMR Experiment | ||||||||
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Experiment | Type | Sample Contents | Solvent | Ionic Strength | pH | Pressure | Temperature (K) | Spectrometer |
1 | 2D NOESY | 0.8mM ZO2PDZ2 domain, 20mM phosphate buffer (pH 6.5), 50mM NaCl, 1mM EDTA, 90% H2O, 10% D2O | 90% H2O/10% D2O | 20mM phosphate buffer, 50mM NaCl | 6.5 | 1 atm | 310 | |
2 | 2D TOCSY | 0.8mM ZO2PDZ2 domain, 20mM phosphate buffer (pH 6.5), 50mM NaCl, 1mM EDTA, 90% H2O, 10% D2O | 90% H2O/10% D2O | 20mM phosphate buffer, 50mM NaCl | 6.5 | 1 atm | 310 | |
3 | 3D_15N-separated_NOESY | 0.8mM 15N, 13C-labeled ZO2PDZ2 domain, 20mM phosphate buffer(pH 6.5), 50mM NaCl, 1mM EDTA, 90% H2O, 10% D2O | 90% H2O/10% D2O | 20mM phosphate buffer, 50mM NaCl | 6.5 | 1 atm | 310 | |
4 | 3D_13C-separated_NOESY | 0.8mM 15N, 13C-labeled ZO2PDZ2 domain, 20mM phosphate buffer (pH 6.5), 50mM NaCl, 1mM EDTA | 100% D2O | 20mM phosphate buffer, 50mM NaCl | 6.5 | 1 atm | 310 | |
5 | 3D_13C F1-filtered,F3-edited NOESY | 0.8mM 15N, 13C-labeled ZO2PDZ2 domain, 20mM phosphate buffer (pH 6.5), 50mM NaCl, 1mM EDTA | 100% D2O | 20mM phosphate buffer, 50mM NaCl | 6.5 | 1 atm | 310 |
NMR Spectrometer Information | |||
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Spectrometer | Manufacturer | Model | Field Strength |
1 | Bruker | DMX | 600 |
NMR Refinement | ||
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Method | Details | Software |
distance geometry, simulated annealing, molecular dynamics | the structures are based on a total of 3076 restraints, 2916 are NOE-derived distance constraints, 160 dihedral angle restraints,76 distance restraints from hydrogen bonds. | CNS |
NMR Ensemble Information | |
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Conformer Selection Criteria | structures with the lowest energy |
Conformers Calculated Total Number | 200 |
Conformers Submitted Total Number | 20 |
Representative Model | 1 (closest to the average) |
Additional NMR Experimental Information | |
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Details | Intermolecular interactions were identified in a 13C/15N-filtered (F1), 13C-edited(F3) 3D NOESY spectrum |
Computation: NMR Software | ||||
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# | Classification | Version | Software Name | Author |
1 | structure solution | CNS | 1.1 | A.T. Brunger |
2 | processing | NMRPipe | 2.2 | F. Delaglio and A. Bax |
3 | data analysis | MOLMOL | 2k.2 | Koradi |
4 | data analysis | TALOS | Cornilescu | |
5 | data analysis | Sparky | 3 | T.D.Goddard and D.G.Kneller |
6 | refinement | CNS | 1.1 | A.T. Brunger |