2AGH

Structural basis for cooperative transcription factor binding to the CBP coactivator


SOLUTION NMR
NMR Experiment
ExperimentTypeSample ContentsSolventIonic StrengthpHPressureTemperature (K)Spectrometer
12D 15N HSQC0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
22D 13C HSQC0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
32D 13C-{13C} SED0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
42D 13C-{15N} SED0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
53D HNCA0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
63D HNCACB0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
73D CBCA(CO)NH0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
83D HBHA(CBCACO)NH0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
93D C(CO)NH-TOCSY0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
103D H(CCO)NH-TOCSY0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
113D CCH-COSY0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
123D CCH-TOCSY0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
133D HCCH-TOCSY0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
143D HACAHB-COSY0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
153D HNHB0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
163D HNHA0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
173D 13C-EDITED NOESY0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
183D 13C,15N-EDITED NOESY0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
193D 12C-FILTERED/13C-EDITED NOESY0.5-0.8 mM 15N,13C-labeled protein (e.g. CBP) + 3 fold excess of unlabeled binding partners (MYB + MLL)20 mM Tris-d3-acetate, 50 mM NaCl, 0.2% sodium azide, 10% D2O5.51 atm298
NMR Spectrometer Information
SpectrometerManufacturerModelField Strength
1BrukerAVANCE900
2BrukerAVANCE800
3BrukerDRX750
4BrukerDRX600
NMR Refinement
MethodDetailsSoftware
simulated annealingiNITIAL STRUCTURE CALCULATION USING TORSION ANGLE DYNAMICS IN DYANA FOLLOWED BY REFINEMENT IN AMBER USING SIMULATED ANNEALING AND ENERGY MINIMIZATION IN VACUO AND GENERALIZED BORN POTENTIALAmber
NMR Ensemble Information
Conformer Selection Criterialowest energy and lowest distance and angle violations
Conformers Calculated Total Number100
Conformers Submitted Total Number20
Representative Model1 (lowest energy)
Computation: NMR Software
#ClassificationVersionSoftware NameAuthor
1refinementAmberDavid Case et al.
2structure calculationDYANAGuntert, Braun and Wuthrich