Experiment: 5M8I

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.6 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 1.8 mM unlabeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
2	2D 1H-13C HSQC aliphatic	0.6 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 1.8 mM unlabeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
3	2D 1H-13C HSQC aromatic	0.6 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 1.8 mM unlabeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
4	3D 1H-15N NOESY	0.6 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 1.8 mM unlabeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
5	3D 1H-13C NOESY aliphatic	0.6 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 1.8 mM unlabeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
14	3D F1 13C-filtered 1H-1H NOESY 13C-1H HSQC aromatic	0.8 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 0.8 mM non-labeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 750
13	3D F1 13C-filtered 1H-1H NOESY 13C-1H HSQC aliphatic	0.8 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 0.8 mM non-labeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
12	natural abundance 2D 13C-1H HSQC	0.8 mM [U-99% 15N] CUG-BP2 third RNA Recognition Motif, 0.8 mM non-labeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
11	2D F2 13C-filtered 1H-1H NOESY	0.8 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 0.8 mM non-labeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
10	2D 1H-1H NOESY	0.8 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 0.8 mM non-labeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
9	2D 1H-1H TOCSY	0.8 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 0.8 mM non-labeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
8	3D HNHA	0.6 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 1.8 mM unlabeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700
7	2D 1H-15N HSQC long range	0.6 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 1.8 mM unlabeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 600
6	3D 1H-13C NOESY aromatic	0.6 mM [U-99% 13C; U-99% 15N] CUG-BP2 third RNA Recognition Motif, 1.8 mM unlabeled 5'-UUUAA-3' RNA	95% H2O/5% D2O	130 mM	5.8	ambient Pa	298	Bruker AVANCE III 700

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE III	600
5	Bruker	AVANCE III	700
4	Bruker	AVANCE III	750
3	Bruker	AVANCE III	900
2	Bruker	AVANCE III HD	600

NMR Refinement
Method	Details	Software
simulated annealing		Amber

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy and least restraints violation
Conformers Calculated Total Number	50
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin		Bruker Biospin
2	processing	TopSpin		Bruker Biospin
3	chemical shift assignment	Sparky		Goddard
4	data analysis	Sparky		Goddard
5	peak picking	CANDID		Herrmann, Guntert and Wuthrich
6	structure calculation	CYANA		Guntert, Mumenthaler and Wuthrich
7	refinement	Amber		Case, Darden, Cheatham III, Simmerling, Wang, Duke, Luo, ... and Kollman

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.