6I3R

Structure, dynamics and roX2-lncRNA binding of tandem double-stranded RNA binding domains dsRBD1/2 of Drosophila helicase MLE

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 800
2	3D HCCH-TOCSY	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 800
3	3D HNCACB	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 800
4	3D HNCA	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 800
5	3D CBCA(CO)NH	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 800
6	3D 1H-13C NOESY aliphatic	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 600
7	3D H(CCO)NH	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 600
8	3D HNCO	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 600
9	3D HBHA(CO)NH	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 600
10	3D 1H-15N NOESY	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 600
11	2D 1H-13C HSQC aromatic	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 800
12	3D 1H-13C NOESY aromatic	0.5 mM [U-13C; U-15N] MLE dsRBD1,2	90% H2O/10% D2O	200 mM	6	1 bar	298	Bruker AVANCE III 600

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE III	600
3	Bruker	AVANCE III	800

NMR Refinement
Method	Details	Software
simulated annealing		ARIA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	10
Conformers Submitted Total Number	10
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	chemical shift assignment	CcpNmr Analysis		CCPN
2	structure calculation	CYANA		Guntert, Mumenthaler and Wuthrich
3	refinement	ARIA		Linge, O'Donoghue and Nilges

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.