1O0P

Solution Structure of the third RNA Recognition Motif (RRM) of U2AF65 in complex with an N-terminal SF1 peptide

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_13C-separated_NOESY	1mM 15N,13C U2AF65-RRM3 + 1mM unlabeled SF1_10-25, 30mM phosphate buffer, 20mM NaCL, 3mM DTT	100% D2O	50mM salt	6.4	ambient	295
2	3D_15N-separated_NOESY	1mM 15N,13C U2AF65-RRM3 + 1mM unlabeled SF1_10-25, 30mM phosphate buffer, 20mM NaCL, 3mM DTT	90% H2O/10% D2O	50mM salt	6.4	ambient	295
3	3D_13C/15N-edited/filtered_NOESY	1mM 15N,13C U2AF65-RRM3 + 1mM unlabeled SF1_10-25, 30mM phosphate buffer, 20mM NaCL, 3mM DTT	100% D2O	50mM salt	6.4	ambient	295

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

NMR Refinement
Method	Details	Software
Restrained molecular dynamics using CNS and ARIA for ambiguous distance restraints	THE EXPERIMENTALLY DETERMINED DISTANCE (3232 NOEs, INCLUDING 258 FOR THE SF1 PEPTIDE, 64 INTERMOLECULAR, AND 2*35 FOR HYDROGEN BONDS) AND DIHEDRAL ANGLE RESTRAINTS (99) WERE APPLIED IN A MIXED TORSION AND CARTESIAN ANGLE DYNAMICS/SIMULATED ANNEALING PROTOCOL. STRUCTURAL QUALITY WAS ASSESED WITH PROCHECK.	NMRPipe

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations,structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	NMRPipe	2.1	DELAGIO
2	data analysis	XEASY		BARTELS
3	structure solution	ARIA	1.2	NILGES
4	structure solution	CNS	1.1	BRUNGER
5	refinement	CNS	1.1	BRUNGER

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.