Experiment: 5HOU

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D 1H-15N NOESY	1.0 mM [U-15N] p53TAD - [NA] TAZ1 p53TAD-TAZ1 fusion protein, 20 mM TRIS, 50 mM sodium chloride, 2 mM DTT, 5 % [U-2H] D2O, 95% H2O, 5% D2O	95% H2O/5% D2O	50 mM	6.8	1 atm	308	Bruker DRX 800
2	3D 1H-13C NOESY	1.0 mM [U-15N][U-13C] p53TAD - [NA] TAZ1 p53TAD-TAZ1 fusion protein, 20 mM [U-2H] TRIS, 50 mM sodium chloride, 2 mM [U-2H] DTT, 99 % [U-2H] D2O	99% D2O, 1% H2O	50 mM	6.4	1 atm	308	Bruker DRX 800
3	3D 1H-15N NOESY	1.0 mM [NA] p53TAD - [U-15N] TAZ1 p53TAD-TAZ1 fusion protein, 20 mM TRIS, 50 mM sodium chloride, 2 mM DTT, 5 % [U-2H] D2O	95% H2O/5% D2O	50 mM	6.8	1 atm	308	Bruker DRX 800
4	3D 1H-13C NOESY	1.0 mM [NA] p53TAD - [U-15N][U-13C] TAZ1 p53TAD-TAZ1 fusion protein, 20 mM [U-2H] TRIS, 50 mM sodium chloride, 2 mM [U-2H] DTT, 99 % [U-2H] D2O	99% D2O, 1% H2O	50 mM	6.4	1 atm	308	Bruker DRX 800
5	3D HNCACB	1.0 mM [U-15N][U-13C] p53TAD - [NA] TAZ1 p53TAD-TAZ1 fusion protein, 20 mM TRIS, 50 mM sodium chloride, 2 mM DTT, 5 % [U-2H] D2O	95% H2O/5% D2O	50 mM	6.8	1 atm	308	Bruker DRX 600
6	3D HN(CO)CA	1.0 mM [U-15N][U-13C] p53TAD - [NA] TAZ1 p53TAD-TAZ1 fusion protein, 20 mM TRIS, 50 mM sodium chloride, 2 mM DTT, 5 % [U-2H] D2O	95% H2O/5% D2O	50 mM	6.8	1 atm	308	Bruker DRX 600
7	3D HNCACB	1 mM [NA] p53TAD - [U-15N][U-13C] TAZ1 p53TAD-TAZ1 fusion protein, 20 mM TRIS, 50 mM sodium chloride, 2 mM DTT, 5 % [U-2H] D2O	95% H2O/5% D2O	50 mM	6.8	1 atm	308	Bruker DRX 600
8	3D HN(CO)CA	1 mM [NA] p53TAD - [U-15N][U-13C] TAZ1 p53TAD-TAZ1 fusion protein, 20 mM TRIS, 50 mM sodium chloride, 2 mM DTT, 5 % [U-2H] D2O	95% H2O/5% D2O	50 mM	6.8	1 atm	308	Bruker DRX 600
9	3D HCCH-TOCSY	1.0 mM [U-15N][U-13C] p53TAD - [NA] TAZ1 p53TAD-TAZ1 fusion protein, 20 mM TRIS, 50 mM sodium chloride, 2 mM DTT, 5 % [U-2H] D2O	95% H2O/5% D2O	50 mM	6.4	1 atm	308	Bruker DRX 600
10	3D HCCH-TOCSY	1 mM [NA] p53TAD - [U-15N][U-13C] TAZ1 p53TAD-TAZ1 fusion protein, 20 mM TRIS, 50 mM sodium chloride, 2 mM DTT, 5 % [U-2H] D2O	95% H2O/5% D2O	50 mM	6.4	1 atm	308	Bruker DRX 600

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	800
2	Bruker	DRX	600

NMR Refinement
Method	Details	Software
simulated annealing, molecular dynamics	Initial structure generation, Refinement of CYANA generated structures	NMRView

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	chemical shift assignment	NMRView	5.0.04	Johnson, One Moon Scientific
2	structure calculation	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
3	refinement	Amber	12	Case, Darden, Cheatham III, Simmerling, Wang, Duke, Luo, and Kollman
4	data analysis	PSVS	1.5	Bhattacharya and Montelione

RCSB PDB Core Operations are funded by the U.S. National Science Foundation (DBI-2321666), 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.