1QU5

NMR STRUCTURE OF A NEW PHOSPHOTYROSINE BINDING DOMAIN CONTAINING THE FHA2 DOMAIN OF RAD 53

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_13C-SEPARATED_NOESY	0.2~0.5 MM FHA2, U-15N, 13C; 100 MM SODIUM PHOSPHATE, 1MM DTT, 1MM EDTA		0.2	6.5	AMBIENT	293
2	3D_15N-SEPARATED_NOESY	0.2~0.5 MM FHA2, U-15N, 13C; 100 MM SODIUM PHOSPHATE, 1MM DTT, 1MM EDTA		0.2	6.5	AMBIENT	293
3	4D_13C-SEPARATED_NOESY	0.2~0.5 MM FHA2, U-15N, 13C; 100 MM SODIUM PHOSPHATE, 1MM DTT, 1MM EDTA		0.2	6.5	AMBIENT	293
4	HCCH-TOCSY	0.2~0.5 MM FHA2, U-15N, 13C; 100 MM SODIUM PHOSPHATE, 1MM DTT, 1MM EDTA		0.2	6.5	AMBIENT	293
5	15N-1H-TROSY-HN(CO)CACB	0.2~0.5 MM FHA2, U-15N, 13C, 2H (~70%) TRIPLE LABELED; 100 MM PHOSPHATE, 1MM DTT, 1MM EDTA		0.2	6.5	AMBIENT	293
6	15N-1H-TROSY-HNCACB	0.2~0.5 MM FHA2, U-15N, 13C, 2H (~70%) TRIPLE LABELED; 100 MM PHOSPHATE, 1MM DTT, 1MM EDTA		0.2	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, MOLECULAR DYNAMICS	IN THE FIRST STAGE, THE SIMULATED ANNEALING STRUCTURES WERE DETERMINED BASED ON THE EXPERIMENTAL INTER PROTON DISTANCE RESTRAINTS (2651 IN TOTAL). THE RESULTING STRUCTURES WERE THEN USED AS INITIAL STRCUTURES FOR THE SECOND STAGE OF SIMULATED ANNEALING CALCULATIONS WHERE, IN ADDITION TO THE DISTANCE RESTRAINTS, THE STRUCTURES WERE REFINED AGAINST SECONDARY 13C(ALPHA)/13C(BETA) CHEMICAL SHIFT RESTRAINTS.	X-PLOR

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	32
Conformers Submitted Total Number	16
Representative Model	1 (minimized average structure)

Additional NMR Experimental Information
Details	THE STRUCTURAL DETERMINATION WAS BENEFITED BY THE TROSY TYPE EXPERIMENTS ON THE TRIPLE-LABLED (13C, 15N, 2H) SAMPLE.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	X-PLOR	3.1	BRUNGER, A. T.
2	structure solution	XwinNMR	2.1	BRUKER
3	structure solution	Felix	98	MSI

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.