2FF0

Solution Structure of Steroidogenic Factor 1 DNA Binding Domain Bound to its Target Sequence in the Inhibin alpha-subunit Promoter

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_15N-separated_NOESY	U-15N-steroidogenic factor 1 DBD:NA-duplex DNA complex	90% H2O/10% D2O	20 mM Tris acetate, 50 uM ZnCl2, 2 mM DTT, 1% glycerol	6.0	ambient	308
2	3D_aliphatic_13C-separated_NOESY	U-15N,U-13C-steroidogenic factor 1 DBD:NA-duplex DNA complex	90% H2O/10% D2O	20 mM Tris acetate, 50 uM ZnCl2, 2 mM DTT, 1% glycerol	6.0	ambient	308
3	3D_aromatic_13C-separated_NOESY	U-15N,U-13C-steroidogenic factor 1 DBD:NA-duplex DNA complex	90% H2O/10% D2O	20 mM Tris acetate, 50 uM ZnCl2, 2 mM DTT, 1% glycerol	6.0	ambient	308
4	2D_15N,13C-double-half-filtered_NOESY	U-15N,U-13C-steroidogenic factor 1 DBD:NA-duplex DNA complex	90% H2O/10% D2O	20 mM Tris acetate, 50 uM ZnCl2, 2 mM DTT, 1% glycerol	6.0	ambient	308
5	2D_15N,13C-double-half-filtered_NOESY	U-15N,U-13C-steroidogenic factor 1 DBD:NA-duplex DNA complex	100% D2O	20 mM Tris acetate, 50 uM ZnCl2, 2 mM DTT, 1% glycerol	6.0	ambient	308
6	3D_13C-filtered_13C-edited_NOESY	U-15N,U-13C-steroidogenic factor 1 DBD:NA-duplex DNA complex	90% H2O/10% D2O	20 mM Tris acetate, 50 uM ZnCl2, 2 mM DTT, 1% glycerol	6.0	ambient	308

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600

NMR Refinement
Method	Details	Software
Simulated annealing and cartesian molecular dynamics		Felix

NMR Ensemble Information
Conformer Selection Criteria	lowest restraint energies, restraint violations, and RMS deviations from ideal covalent geometry
Conformers Calculated Total Number	200
Conformers Submitted Total Number	16
Representative Model	1 (best captures the intermolecular interactions observed in the ensemble)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	Felix	98.0	Accelrys Inc
2	refinement	ARIA	1.2	Nilges et al.
3	refinement	CNS	1.1	Brunger et al.
4	collection	VNMR	6.1	Varian Inc

RCSB PDB Core Operations are funded by the National Science Foundation (DBI-1832184), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM133198.