2KGP

Structural basis for stabilization of the tau pre-mRNA splicing regulatory element by Novantrone (Mitoxantrone)

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-1H NOESY	0.8~1.8 mM tau pre-mRNA splicing regulatory element-1	95% H2O/5% D2O	10	6	ambient	277
2	2D 1H-1H TOCSY	0.8~1.8 mM tau pre-mRNA splicing regulatory element-2	100% D2O	10	6	ambient	277
3	2D 1H-1H NOESY	0.8~1.8 mM tau pre-mRNA splicing regulatory element-2	100% D2O	10	6	ambient	277
4	2D 1H-1H TOCSY	0.8 mM [U-99% 13C; U-99% 15N] tau pre-mRNA splicing regulatory element-3	100% D2O	10	6	ambient	277
5	2D 1H-1H NOESY	0.8 mM [U-99% 13C; U-99% 15N] tau pre-mRNA splicing regulatory element-3	100% D2O	10	6	ambient	277
6	2D 1H-13C HSQC	0.8~1.2 mM Partial deuterated nucleotides tau pre-mRNA splicing regulatory element-4	100% D2O	10	6	ambient	277
7	3D 1H-13C NOESY	0.8~1.2 mM Partial deuterated nucleotides tau pre-mRNA splicing regulatory element-4	100% D2O	10	6	ambient	277

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	600
2	Bruker	AVANCE	500
3	Varian	Uniform NMR System	800
4	Bruker	AVANCE	750

NMR Refinement
Method	Details	Software
simulated annealing		NMRPipe

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	chemical shift assignment	Sparky		Goddard
3	chemical shift calculation	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
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 R01GM133198.