1XSU

Solution structure of E.coli RNase P RNA P4 stem, U69C/C70U mutation, complexed with cobalt (III) hexammine.

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D NOESY	2mM P4 U69C/C70U RNA; 100mM NaCl; 10mM phosphate buffer; 90% H2O, 10% D2O	90% H2O/10% D2O	100 mM NaCl	6.4	ambient	288
2	2D NOESY	2mM P4 U69C/C70U RNA; 100mM NaCl; 10mM phosphate buffer; 100% D2O	100% D2O	100 mM NaCl	6.4	ambient	288
3	2D NOESY	2mM P4 U69C/C70U RNA; 100mM NaCl; 6mM hexammine cobalt chloride; 10mM phosphate buffer; 90% H2O, 10% D2O	90% H2O/10% D2O	100 mM NaCl, 6 mM Co(NH3)6Cl	6.4	ambient	288
4	DQF-COSY	2mM P4 U69C/C70U RNA; 100mM NaCl; 10mM phosphate buffer; 100% D2O	100% D2O	100 mM NaCl	6.4	ambient	288
5	31P-1H-Hetero-COSY	2mM P4 U69C/C70U RNA; 100mM NaCl; 10mM phosphate buffer; 100% D2O	100% D2O	100 mM NaCl	6.4	ambient	288
6	31P-1H-Hetero-TOCSY-NOESY	2mM P4 U69C/C70U RNA; 100mM NaCl; 10mM phosphate buffer; 100% D2O	100% D2O	100 mM NaCl	6.4	ambient	288

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DMX	600

NMR Refinement
Method	Details	Software
restrained molecular dynamics and simulated annealing	The average structure is based on the superposition of 18 structures after refinement. The average RMS deviation between the ensemble and the average structure is 1.87 Angstrom. A total of 261 NOE-derived distance constraints, 246 dihedral constraints and 48 distance constraints from hydrogen bonds were used in refinement. 12 NOE derived intermolecular distance constraints were used to localize the bound cobalt (III) hexammine.	XwinNMR

NMR Ensemble Information
Conformer Selection Criteria
Conformers Calculated Total Number
Conformers Submitted Total Number	1
Representative Model	1 (minimized average structure)

Additional NMR Experimental Information
Details	The structure was determined using standard 2D homonuclear techniques as well as 13C and 31P heteronuclear experiments performed at natural abundance. Intermolecular NOE crosspeaks between RNA protons and cobalt (III) hexammine protons and intermolecular distance constraints derived thereof were used to determine the site of cobalt (III) hexammine binding.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	XwinNMR	1.2	Bruker
2	processing	NMRPipe	2.1	F. Delaglio
3	structure solution	X-PLOR	3.1	A. Bruenger
4	refinement	X-PLOR	3.1	A. Bruenger

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.