2A0T

NMR structure of the FHA1 domain of Rad53 in complex with a biological relevant phosphopeptide derived from Madt1

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D 13C-edited 13C/15N-filtered NOESY	0.5 mM FHA1 U-13C, 15N, ~0.8 mM peptide, 10 mM sodium phosphate buffer (pH 6.5), 1 mM DTT,1 mM EDTA, 90% H2O, 10% D2O	90% H2O/10% D2O	10 mM sodium phosphate, 1 mM DTT, and 1 mM EDTA	6.5	ambient	293
2	3D 13C/15N-filtered, 13C-edited NOESY	0.5 mM FHA1 U-13C, 15N, ~0.8 mM peptide, 10 mM sodium phosphate buffer (pH 6.5), 1 mM DTT, 1 mM EDTA, 100% D2O	100% D2O	10 mM sodium phosphate, 1 mM DTT, and 1 mM EDTA	6.5	ambient	293
3	3D_13C-separated_NOESY	0.5 mM FHA1 U-13C, 15N, ~0.8 mM peptide, 10 mM sodium phosphate buffer (pH 6.5), 1 mM DTT, 1 mM EDTA, 100% D2O	100% D2O	10 mM sodium phosphate, 1 mM DTT, and 1 mM EDTA	6.5	ambient	293
4	2D 13C/15N-filtered NOESY	0.5 mM FHA1 U-13C, 15N, ~0.8 mM peptide, 10 mM sodium phosphate buffer (pH 6.5), 1 mM DTT,1 mM EDTA, 90% H2O, 10% D2O	90% H2O/10% D2O	10 mM sodium phosphate, 1 mM DTT, and 1 mM EDTA	6.5	ambient	293
5	2D 13C/15N-filtered TOCSY	0.5 mM FHA1 U-13C, 15N, ~0.8 mM peptide, 10 mM sodium phosphate buffer (pH 6.5), 1 mM DTT, 1 mM EDTA, 100% D2O	100% D2O	10 mM sodium phosphate, 1 mM DTT, and 1 mM EDTA	6.5	ambient	293
6	2D 13C/15N-filtered COSY	0.5 mM FHA1 U-13C, 15N, ~0.8 mM peptide, 10 mM sodium phosphate buffer (pH 6.5), 1 mM DTT,1 mM EDTA, 90% H2O, 10% D2O	90% H2O/10% D2O	10 mM sodium phosphate, 1 mM DTT, and 1 mM EDTA	6.5	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	800
2	Bruker	DMX	600
3	Bruker	AVANCE	800
4	Bruker	AVANCE	500

NMR Refinement
Method	Details	Software
simulated annealing	The complex structures are generated using a total of 2524 NMR constraints including constraints in this PDB file and constraints of free FHA1 in PDB access code 1K3J	XwinNMR

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	50
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average, lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	XwinNMR	3.5	Bruker
2	processing	NMRPipe		Delaglio, F.
3	data analysis	NMRView	5	Johnson, B.
4	structure solution	CNS	1.1	Brunger et al.
5	refinement	CNS	1.1	Brunger 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.