1JU7

NMR Solution Structure of the RNA Hairpin Binding Site for the Histone Stem-loop Binding Protein

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	NOESY	2.8 mM unlabeled 20 mM KPi, 20 mM KCl, 0.02 mM EDTA, pH 6.8	100% D2O	100 mM	6.8	ambient	301
2	3D_13C-separated_NOESY	2.3 mM U-13C; 20 mM KPi, 20 mM KCl, 0.02 mM EDTA, pH 6.8	100% D2O	100 mM	6.8	ambient	301
3	3D_15N-separated_NOESY	2.5 mM U-15N; 20 mM KPi, 20 mM KCl, 0.02 mM EDTA, pH 6.8	10% D2O, 90% H2O	100 mM	6.8	ambient	280
4	3D_13C-separated_NOESY	2.0 mM U-13C,C5-2H; 20 mM KPi, 20 mM KCl, 0.02 mM EDTA, pH 6.8	100% D2O	100 mM	6.8	ambient	301
5	DQF-COSY	2.5 mM U-15N; 20 mM KPi, 20 mM KCl, 0.02 mM EDTA, pH 6.8	10% D2O, 90% H2O	100 mM	6.8	ambient	301
6	HetCor	2.8 mM unlabeled 20 mM KPi, 20 mM KCl, 0.02 mM EDTA, pH 6.8	100% D2O	100 mM	6.8	ambient	301

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AMX	500

NMR Refinement
Method	Details	Software
simulated annealing and molecular dynamics	Calculations were performed using 232 conformationally restrictive NOE derived distance constraints and 55 backbone and 15 ribose torsion angle constraints. Base pair constraints were introduced for six base pairs using heavy atom-heavy atom constraints. Coordinates are for the hairpin domain only. 5' flanking (ggccaaa) and 3' flanking (accca) coordinates are not included with this deposition. Few constraints were obtained for these very dynamic regions and although they were included as part of the structure calculation, their conformations appear to be randomly distributed.	Felix

NMR Ensemble Information
Conformer Selection Criteria	structures with acceptable covalent geometry, structures with the least restraint violations, structures with the lowest energy
Conformers Calculated Total Number	60
Conformers Submitted Total Number	10
Representative Model	5 (closest to the average)

Additional NMR Experimental Information
Details	This structure was determined using a variety of 2D and 3D homo- and hetero-nuclear NMR methods.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	Felix	980	MSI-Biosym
2	refinement	X-PLOR	3.851	Brunger, A.

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.