1F7I

SOLUTION STRUCTURE OF THE RNASE P RNA (M1 RNA) P4 STEM C70U MUTANT OLIGORIBONUCLEOTIDE COMPLEXED WITH COBALT (III) HEXAMINE ,NMR, ENSEMBLE OF 12 STRUCTURES

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D NOESY	2 mM P4m RNA; 100 mM sodium chloride; 10 mM phosphate buffer; 90% H2O, 10% D2O	90% H2O/10% D2O	100 mM Na	6.5	ambient	288
2	2D NOESY	2 mM P4m RNA; 100 mM sodium chloride; 10 mM phosphate buffer; 99.96% D2O	99.96% D2O	100 mM Na	6.5	ambient	288
3	2D NOESY	2 mM P4m RNA; 100 mM sodium chloride; 10 mM magnesium chloride; 10 mM phosphate buffer; 90% H2O, 10% D2O	90% H2O/10% D2O	100 mM Na, 10 mM Mg	6.5	ambient	288
4	2D NOESY	2 mM P4m RNA; 100 mM sodium chloride; 3 mM hexammine cobalt chloride; 10 mM phosphate buffer; 90% H2O, 10% D2O	90% H2O/10% D2O	100 mM Na, 3 mM Co(NH3)6	6.5	ambient	288
5	DQF-COSY	2 mM P4m RNA; 100 mM sodium chloride; 10 mM phosphate buffer; 99.96% D2O	99.96% D2O	100 mM Na	6.5	ambient	288
6	31P-1H-COSY	2 mM P4m RNA; 100 mM sodium chloride; 10 mM phosphate buffer; 99.96% D2O	99.96% D2O	100 mM Na	6.5	ambient	288
7	13C-1H-HMQC	2 mM P4m RNA; 100 mM sodium chloride; 10 mM phosphate buffer; 99.96% D2O	99.96% D2O	100 mM Na	6.5	ambient	288

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

NMR Refinement
Method	Details	Software
restrained molecular dynamics; simulated annealing	The structures are based on a total of 268 NOE derived distance constraints, 171 dihedral restraints and 49 distance restraints from hydrogen bonds. 12 NOE derived intermolecular distance constraints were used to localize the bound cobalt(III) hexammine. The 12 structures with lowest NOE and dihedral angle violation energies are presented	XwinNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	50
Conformers Submitted Total Number	12
Representative Model	7 (closest to the average)

Additional NMR Experimental Information
Details	This structure was determined using standard 2D homonuclear techniques and 13C and 31P heteronuclear techniques at natural abundance. Distance constraints derived from intermolecular NOE crosspeaks between RNA protons and cobalt (III) hexammine protons were used to determine the site of cobalt (III) hexammine binding in the complex structure

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	XwinNMR	3.1	Bruker
2	processing	Felix	95	MSI
3	structure solution	X-PLOR	3.841	Brunger
4	refinement	X-PLOR	3.841	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.