Experiment: 2RT3

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	50 mM potassium phosphate-6, 100 mM sodium chloride-7, 1 mM TCEP-8, 0.1 mM EDTA-9, 1.2 mM [U-99% 15N] Nrd1 RRM2-10	93% H2O/7% D2O		6.0	ambient	303
2	3D HNCACB	50 mM potassium phosphate-1, 100 mM sodium chloride-2, 1 mM TCEP-3, 0.1 mM EDTA-4, 1.0 mM [U-99% 13C; U-99% 15N] Nrd1 RRM2-5	93% H2O/7% D2O		6.0	ambient	303
3	3D CBCA(CO)NH	50 mM potassium phosphate-1, 100 mM sodium chloride-2, 1 mM TCEP-3, 0.1 mM EDTA-4, 1.0 mM [U-99% 13C; U-99% 15N] Nrd1 RRM2-5	93% H2O/7% D2O		6.0	ambient	303
4	3D HN(CA)CO	50 mM potassium phosphate-1, 100 mM sodium chloride-2, 1 mM TCEP-3, 0.1 mM EDTA-4, 1.0 mM [U-99% 13C; U-99% 15N] Nrd1 RRM2-5	93% H2O/7% D2O		6.0	ambient	303
5	3D HNCO	50 mM potassium phosphate-1, 100 mM sodium chloride-2, 1 mM TCEP-3, 0.1 mM EDTA-4, 1.0 mM [U-99% 13C; U-99% 15N] Nrd1 RRM2-5	93% H2O/7% D2O		6.0	ambient	303
6	3D H(CCO)NH	50 mM potassium phosphate-1, 100 mM sodium chloride-2, 1 mM TCEP-3, 0.1 mM EDTA-4, 1.0 mM [U-99% 13C; U-99% 15N] Nrd1 RRM2-5	93% H2O/7% D2O		6.0	ambient	303
7	3D C(CO)NH	50 mM potassium phosphate-1, 100 mM sodium chloride-2, 1 mM TCEP-3, 0.1 mM EDTA-4, 1.0 mM [U-99% 13C; U-99% 15N] Nrd1 RRM2-5	93% H2O/7% D2O		6.0	ambient	303
8	4D H(CCO)NH	50 mM potassium phosphate-1, 100 mM sodium chloride-2, 1 mM TCEP-3, 0.1 mM EDTA-4, 1.0 mM [U-99% 13C; U-99% 15N] Nrd1 RRM2-5	93% H2O/7% D2O		6.0	ambient	303
9	3D 1H-13C NOESY	50 mM potassium phosphate-11, 100 mM sodium chloride-12, 1 mM TCEP-13, 0.1 mM EDTA-14, 1.0 mM [U-99% 13C; U-99% 15N] Nrd1 RRM2-15	100% D2O		6.0	ambient	303
10	3D 1H-15N NOESY	50 mM potassium phosphate-6, 100 mM sodium chloride-7, 1 mM TCEP-8, 0.1 mM EDTA-9, 1.2 mM [U-99% 15N] Nrd1 RRM2-10	93% H2O/7% D2O		6.0	ambient	303
11	3D HNHB	50 mM potassium phosphate-6, 100 mM sodium chloride-7, 1 mM TCEP-8, 0.1 mM EDTA-9, 1.2 mM [U-99% 15N] Nrd1 RRM2-10	93% H2O/7% D2O		6.0	ambient	303
12	3D HA(CA)HB	50 mM potassium phosphate-11, 100 mM sodium chloride-12, 1 mM TCEP-13, 0.1 mM EDTA-14, 1.0 mM [U-99% 13C; U-99% 15N] Nrd1 RRM2-15	100% D2O		6.0	ambient	303
13	2D 1H-13C CT-HSQC	50 mM potassium phosphate-16, 100 mM sodium chloride-17, 1 mM TCEP-18, 0.1 mM EDTA-19, 0.35 mM [U-10% 13C; U-99% 15N] Nrd1 RRM2-20	93% H2O/7% D2O		6.0	ambient	303

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

NMR Refinement
Method	Details	Software
simulated annealing		CNS

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
2	structure solution	CYANA		Guntert, Mumenthaler and Wuthrich
3	chemical shift assignment	Sparky		Goddard
4	data analysis	TALOS		Cornilescu, Delaglio and Bax
5	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
6	collection	TopSpin		Bruker Biospin

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.