2JQI

NMR Structure of the Rad53 FHA1 domain in complex with a phosphothreonien peptide derived from Rad53 SCD1

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D 1H-13C NOESY	0.5 mM [U-13C; U-15N] protein, 0.8 mM peptide, 10 mM sodium phosphate, 1 mM DTT, 1 mM EDTA	100% D2O	10 mM sodium phosphate	6.5	ambient	293
2	3D 13C/15N-filtered (f1), 13C-edited (f3) NOESY	0.5 mM [U-13C; U-15N] protein, 0.8 mM peptide, 10 mM sodium phosphate, 1 mM DTT, 1 mM EDTA	90% H2O/10% D2O	10 mM sodium phosphate	6.5	ambient	293
3	3D 13C/15N-filtered (f1), 13C-edited (f3) NOESY	0.5 mM [U-13C; U-15N] protein, 0.8 mM peptide, 10 mM sodium phosphate, 1 mM DTT, 1 mM EDTA	100% D2O	10 mM sodium phosphate	6.5	ambient	293
4	3D 13C-edited (f1), 13C/15N-filtered (f3) NOESY	0.5 mM [U-13C; U-15N] protein, 0.8 mM peptide, 10 mM sodium phosphate, 1 mM DTT, 1 mM EDTA	100% D2O	10 mM sodium phosphate	6.5	ambient	293
5	2D 13C/15N-filtered (f1,f2) NOESY	0.5 mM [U-13C; U-15N] protein, 0.8 mM peptide, 10 mM sodium phosphate, 1 mM DTT, 1 mM EDTA	90% H2O/10% D2O	10 mM sodium phosphate	6.5	ambient	293
6	2D 13C/15N-filtered (f1,f2) NOESY	0.5 mM [U-13C; U-15N] protein, 0.8 mM peptide, 10 mM sodium phosphate, 1 mM DTT, 1 mM EDTA	100% D2O	10 mM sodium phosphate	6.5	ambient	293
7	2D 13C/15N-filtered (f1,f2) TOCSY	0.5 mM [U-13C; U-15N] protein, 0.8 mM peptide, 10 mM sodium phosphate, 1 mM DTT, 1 mM EDTA	90% H2O/10% D2O	10 mM sodium phosphate	6.5	ambient	293
8	2D 13C/15N-filtered (f1,f2) TOCSY	0.5 mM [U-13C; U-15N] protein, 0.8 mM peptide, 10 mM sodium phosphate, 1 mM DTT, 1 mM EDTA	100% D2O	10 mM sodium phosphate	6.5	ambient	293
9	2D 13C/15N-filtered (f1,f2) COSY	0.5 mM [U-13C; U-15N] protein, 0.8 mM peptide, 10 mM sodium phosphate, 1 mM DTT, 1 mM EDTA	100% D2O	10 mM sodium phosphate	6.5	ambient	293

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

NMR Refinement
Method	Details	Software
simulated annealing		CNS

NMR Ensemble Information
Conformer Selection Criteria	structures with acceptable covalent geometry,structures with the least restraint violations,structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CNS	1.1	Brunger, A.T. et al.
2	processing	NMRPipe		Delaglio, F. et al.
3	data analysis	NMRView		Johnson, B.A. et al.
4	refinement	CNS	1.1	Brunger, A.T. et al.

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.