2JQL

NMR structure of the yeast Dun1 FHA domain in complex with a doubly phosphorylated (pT) 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	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM [U-2H] HEPES, 150 mM NaCl, 1 mM DTT	100% D2O	150 mM NaCl	7.5	ambient	293
2	3D 1H-15N NOESY	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM HEPES, 150 mM NaCl, 1 mM EDTA, 2 mM DTT	90% H2O/10% D2O	150 mM NaCl	7.5	ambient	293
3	3D HNCA	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM HEPES, 150 mM NaCl, 1 mM EDTA, 2 mM DTT	90% H2O/10% D2O	150 mM NaCl	7.5	ambient	293
4	3D HN(CO)CA	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM HEPES, 150 mM NaCl, 1 mM EDTA, 2 mM DTT	90% H2O/10% D2O	150 mM NaCl	7.5	ambient	293
5	3D 13C/15N-filtered (f1),13C-edited (f3) NOESY	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM [U-2H] HEPES, 1 mM DTT, 150 mM NaCl	90% H2O/10% D2O	150 mM NaCl	7.5	ambient	293
6	3D 13C/15N-filtered (f1), 13C-edoted (f3) NOESY	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM [U-2H] HEPES, 150 mM NaCl, 1 mM DTT	100% D2O	150 mM NaCl	7.5	ambient	293
7	3D 13C-edited (f1), 13C/15N-filtered (f3) NOESY	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM [U-2H] HEPES, 150 mM NaCl, 1 mM DTT	100% D2O	150 mM NaCl	7.5	ambient	293
8	2D 13C/15N-filtered (f1,f2) NOESY	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM [U-2H] HEPES, 1 mM DTT, 150 mM NaCl	90% H2O/10% D2O	150 mM NaCl	7.5	ambient	293
9	2D 13C/15N-filtered (f1,f2) NOESY	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM [U-2H] HEPES, 150 mM NaCl, 1 mM DTT	100% D2O	150 mM NaCl	7.5	ambient	293
10	2D 13C/15N-filtered (f1,f2) TOCSY	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM [U-2H] HEPES, 1 mM DTT, 150 mM NaCl	90% H2O/10% D2O	150 mM NaCl	7.5	ambient	293
11	2D 13C/15N-filtered (f1,f2) TOCSY	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM [U-2H] HEPES, 1 mM DTT, 150 mM NaCl	90% H2O/10% D2O	150 mM NaCl	7.5	ambient	293
12	2D 13C/15N-filtered (f1,f2) COSY	0.4 mM [U-13C; U-15N] protein, 0.5 mM peptide, 5 mM [U-2H] HEPES, 150 mM NaCl, 1 mM DTT	100% D2O	150 mM NaCl	7.5	ambient	293

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

NMR Refinement
Method	Details	Software
simulated annealing		NMRPipe

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
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	processing	NMRPipe		Delaglio, F. et al.
2	data analysis	NMRView		Johnson, B.A. et al.
3	structure solution	CNS		Brunger, A.T. et al.
4	refinement	CNS		Brunger, A.T. et al.

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.