2AFF

The solution structure of the Ki67FHA/hNIFK(226-269)3P complex

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_13C-separated_NOESY	0.9 mM Ki67 FHA U-15N,13C; 1 mM hNIFK(226-269)3P unlabeled 5 mM HEPES, 5 mM DTT, 1 mM EDTA, 150 mM NaCl, pH 7.4	93% H2O/7% D2O	150 mM NaCl	7.4	ambient	293
2	3D_15N-separated_NOESY	0.9 mM Ki67 FHA U-15N,13C; 1 mM hNIFK(226-269)3P unlabeled 5 mM HEPES, 5 mM DTT, 1 mM EDTA, 150 mM NaCl, pH 7.4	93% H2O/7% D2O	150 mM NaCl	7.4	ambient	293
3	3D_13C-separated_NOESY	1 mM Ki67 FHA unlabled; 0.9 mM hNIFK(226-269)3P U-15N,13C 5 mM HEPES, 5 mM DTT, 1 mM EDTA, 150 mM NaCl, pH 7.4	93% H2O/7% D2O	150 mM NaCl	7.4	ambient	293
4	3D_15N-separated_NOESY	1 mM Ki67 FHA unlabled; 0.9 mM hNIFK(226-269)3P U-15N,13C 5 mM HEPES, 5 mM DTT, 1 mM EDTA, 150 mM NaCl, pH 7.4	93% H2O/7% D2O	150 mM NaCl	7.4	ambient	293
5	3D_12C/14N-filtered_13C_separated NOESY	0.9 mM Ki67 FHA U-15N,13C; 1 mM hNIFK(226-269)3P unlabeled 5 mM HEPES, 5 mM DTT, 1 mM EDTA, 150 mM NaCl, pH 7.4	93% H2O/7% D2O	150 mM NaCl	7.4	ambient	293
6	3D_12C/14N-filtered_13C_separated NOESY	1 mM Ki67 FHA unlabled; 0.9 mM hNIFK(226-269)3P U-15N,13C 5 mM HEPES, 5 mM DTT, 1 mM EDTA, 150 mM NaCl, pH 7.4	93% H2O/7% D2O	150 mM NaCl	7.4	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	800
2	Bruker	DMX	750
3	Bruker	AVANCE	600
4	Bruker	DMX	500

NMR Refinement
Method	Details	Software
simulated annealing	the structures are based on a total of 3476 constraints, 3141 are distance, 215 dihedral angle, and 120 N-H residual dipolar coupling constraints	X-PLOR

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations,structures with the lowest energy
Conformers Calculated Total Number	512
Conformers Submitted Total Number	100
Representative Model	1 (closest to the average)

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

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	X-PLOR	NIH version	Brunger
2	refinement	X-PLOR	NIH version	Brunger

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.