2LUP

RDC refined solution structure of double-stranded RNA binding domain of S. cerevisiae RNase III (rnt1p) in complex with the terminal RNA hairpin of snr47 precursor

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% 13C; U-100% 15N] protein, 90 % H2O, 10 % [U-100% 2H] D2O, 20 mM sodium phosphate, 150 mM sodium chloride	90% H2O/10% D2O	150	6.5	ambient	298
2	3D HNCACB	1 mM [U-100% 13C; U-100% 15N] protein, 90 % H2O, 10 % [U-100% 2H] D2O, 20 mM sodium phosphate, 150 mM sodium chloride	90% H2O/10% D2O	150	6.5	ambient	298
3	3D HBHA(CO)NH	1 mM [U-100% 13C; U-100% 15N] protein, 90 % H2O, 10 % [U-100% 2H] D2O, 20 mM sodium phosphate, 150 mM sodium chloride	90% H2O/10% D2O	150	6.5	ambient	298
4	3D HNCO	1 mM [U-100% 13C; U-100% 15N] protein, 90 % H2O, 10 % [U-100% 2H] D2O, 20 mM sodium phosphate, 150 mM sodium chloride	90% H2O/10% D2O	150	6.5	ambient	298
5	3D HCCH-TOCSY	1 mM [U-100% 13C; U-100% 15N] protein, 90 % [U-100% 2H] D2O, 10 % sodium chloride, 150 mM sodium phosphate	100% D2O	150	6.5	ambient	298
6	3D HCCH-COSY	1 mM [U-100% 13C; U-100% 15N] protein, 90 % [U-100% 2H] D2O, 10 % sodium chloride, 150 mM sodium phosphate	100% D2O	150	6.5	ambient	298
7	3D 1H-15N NOESY	1 mM [U-100% 13C; U-100% 15N] protein, 90 % H2O, 10 % [U-100% 2H] D2O, 20 mM sodium phosphate, 150 mM sodium chloride	90% H2O/10% D2O	150	6.5	ambient	298
8	3D 1H-13C NOESY	1 mM [U-100% 13C; U-100% 15N] protein, 90 % [U-100% 2H] D2O, 10 % sodium chloride, 150 mM sodium phosphate	100% D2O	150	6.5	ambient	298

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

NMR Refinement
Method	Details	Software
simulated annealing	THIS STRUCTURE IS AN RDC REFINED STRUCTURE OF PDB ENTRY 1T4L USING THE SAME RESTRAINTS PLUS ADDITIONAL RDCS [81 RDCS (REFINED) VS 43 RDCS (1T4L)].	TopSpin

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	16
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin		Bruker Biospin
2	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	data analysis	NMRView		Johnson, One Moon Scientific
4	chemical shift assignment	NMRView		Johnson, One Moon Scientific
5	data analysis	TALOS		Cornilescu, Delaglio and Bax
6	structure solution	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
7	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore

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.