2GS0

NMR structure of the complex between the PH domain of the Tfb1 subunit from TFIIH and the activation domain of p53

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D 15N-edited NOESY-HSQC	1.0 mM Tfb1 U-15N, 1.0 mM p53 unlabeled, 1.00 mM EDTA, 20 mM phosphate buffer; 90% H2O, 10% D2O	90% H2O/10% D2O	20 mM sodium phosphate	6.5	ambient	300
2	3D 13C-edited HMQC-NOESY	1.0 mM Tfb1 U-15N,13C, 1.0 mM p53 unlabeled, 1.00 mM EDTA, 20 mM phosphate buffer; 100% D20	100% D20	20 mM sodium phosphate	6.5	ambient	300
3	3D 15N-13C {F1}-filtered {F3}-edited NOESY	1.0 mM Tfb1 U-15N,13C, 1.0 mM p53 unlabeled, 1.00 mM EDTA, 20 mM phosphate buffer; 100% D20	100% D20	20 mM sodium phosphate	6.5	ambient	300
4	2D 1H-15N HSQC	1.0 mM Tfb1 U-15N, 1.0 mM p53 unlabeled, 1.00 mM EDTA, 20 mM phosphate buffer; 90% H2O, 10% D2O	90% H2O/10% D2O	20 mM sodium phosphate	6.5	ambient	300
5	3D 15N-edited NOESY-HSQC	1.0 mM p53 U-15N, 1.0 mM Tfb1 unlabeled, 1.00 mM EDTA, 20 mM phosphate buffer; 90% H2O, 10% D2O	90% H2O/10% D2O	20 mM sodium phosphate	6.5	ambient	300
6	3D 13C-edited HMQC-NOESY	1.0 mM p53 U-15N, 1.0 mM Tfb1 unlabeled, 1.00 mM EDTA, 20 mM phosphate buffer; 100% D20	100% D20	20 mM sodium phosphate	6.5	ambient	300
7	3D 15N-13C {F1}-filtered {F3}-edited NOESY	1.0 mM p53 U-15N, 1.0 mM Tfb1 unlabeled, 1.00 mM EDTA, 20 mM phosphate buffer; 100% D20	100% D20	20 mM sodium phosphate	6.5	ambient	300
8	2D 1H-15N HSQC	1.0 mM p53 U-15N, 1.0 mM Tfb1 unlabeled, 1.00 mM EDTA, 20 mM phosphate buffer; 90% H2O, 10% D2O	90% H2O/10% D2O	20 mM sodium phosphate	6.5	ambient	300
9	2D CT-HMQC	1.0 mM Tfb1 U-15N,13C, 1.0 mM p53 unlabeled, 1.00 mM EDTA, 20 mM phosphate buffer; 100% D20	100% D20	20 mM sodium phosphate	6.5	ambient	300
10	2D CT-HMQ	1.0 mM p53 U-15N, 1.0 mM Tfb1 unlabeled, 1.00 mM EDTA, 20 mM phosphate buffer; 100% D20	100% D20	20 mM sodium phosphate	6.5	ambient	300

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	UNITY	500
2	Varian	UNITY	600
3	Varian	UNITY	800

NMR Refinement
Method	Details	Software
Simulated annealing, with a combination of torsion angle and Cartesian dynamics.	The three-dimensional structures of the complex Tfb1/p53 were determined using a set of 1720 NOE-derived distance restraints, 138 backbone dihedral angle (phi and psi) restraints and 26 distance restraints from hydrogen bonds. Because of the absence of medium-range, long-range and intermolecular NOEs involving residues 20-44 and 59-73 of p53 (chain B), these amino acids were not included in the calculations.	VNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	67
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	The structure was determined using triple-resonance NMR spectroscopy.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VNMR	6.1.C	Varian
2	processing	NMRPipe	2.2	F. Delaglio, S. Grzesiek, G.W. Vuister, G. Zhu, J. Pfeifer and A. Bax
3	data analysis	NMRView	5.0.4	B.A. Johnson and R.A. Blevins
4	structure solution	CNS	1.0	A.T.Brunger, P.D.Adams, G.M.Clore, W.L.Delano, P.Gros, R.W.Grosse-Kunstleve, J.-S.Jiang, J.Kuszewski, M.Nilges, N.S.Pannu, R.J.Read, L.M.Rice, T.Simonson, G.L.Warren
5	refinement	CNS	1.0	A.T.Brunger, P.D.Adams, G.M.Clore, W.L.Delano, P.Gros, R.W.Grosse-Kunstleve, J.-S.Jiang, J.Kuszewski, M.Nilges, N.S.Pannu, R.J.Read, L.M.Rice, T.Simonson, G.L.Warren

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.