2MIS | pdb_00002mis

NMR Localization of Divalent Cations at the Active Site of the Neurospora VS Ribozyme Provides Insights Into RNA-Metal Ion Interactions

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1 mM [U-100% 15N] RNA	90% H2O/10% D2O	5 mM Mg2+, 50 mM potassium chloride	6.5	ambient	288
2	2D 1H-15N HSQC NH2 only	1 mM [U-100% 15N] RNA	90% H2O/10% D2O	5 mM Mg2+, 50 mM potassium chloride	6.5	ambient	288
3	2D 1H-13C HSQC	0.9 mM [U-100% 13C; U-100% 15N] RNA	100% D2O	5 mM Mg2+, 50 mM potassium chloride	6.5	ambient	298
4	2D 1H-15N HSQC long-range	0.9 mM [U-100% 13C; U-100% 15N] RNA	90% H2O/10% D2O	5 mM Mg2+, 50 mM potassium chloride	6.5	ambient	288
5	1D 31P	0.2-0.4 mM phosphorothioate RNA	90% H2O/10% D2O	5 mM Mg2+/Cd2+, 50 mM potassium chloride	6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	500
2	Varian	INOVA	600

NMR Refinement
Method	Details	Software
simulated annealing		X-PLOR NIH

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	500
Conformers Submitted Total Number	21
Representative Model	1 (minimized average structure)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
2	data analysis	NMRView		Johnson, One Moon Scientific
3	data analysis	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore

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 R01GM157729.