Experiment: 2LA4

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-1H NOESY	0.3mM PUB1 RRM3-6, 25mM potassium phosphate-7, 25mM sodium chloride-8, 0.5mM DTT-9, 0.01mM DSS-10	90% H2O/10% D2O		7.0	1 atm	298
2	3D 1H-15N NOESY 2D 1H-15N HSQC	0.3mM [U-99% 15N] PUB1 RRM3-1, 25mM potassium phosphate-2, 25mM sodium chloride-3, 0.5mM DTT-4, 0.01mM DSS-5	90% H2O/10% D2O		7.0	1 atm	298
3	3D HNCO	0.3mM [U-99% 13C; U-99% 15N] PUB1 RRM3-16, 25mM potassium phosphate-17, 25mM sodium chloride-18, 0.5mM DTT-19, 0.1mM DSS-20	90% H2O/10% D2O		7.0	1 atm	298
4	3D HN(CA)CO	0.3mM [U-99% 13C; U-99% 15N] PUB1 RRM3-16, 25mM potassium phosphate-17, 25mM sodium chloride-18, 0.5mM DTT-19, 0.1mM DSS-20	90% H2O/10% D2O		7.0	1 atm	298
5	3D HNCA	0.3mM [U-99% 13C; U-99% 15N] PUB1 RRM3-16, 25mM potassium phosphate-17, 25mM sodium chloride-18, 0.5mM DTT-19, 0.1mM DSS-20	90% H2O/10% D2O		7.0	1 atm	298
6	3D HN(CO)CA	0.3mM [U-99% 13C; U-99% 15N] PUB1 RRM3-16, 25mM potassium phosphate-17, 25mM sodium chloride-18, 0.5mM DTT-19, 0.1mM DSS-20	90% H2O/10% D2O		7.0	1 atm	298
7	3D CBCA(CO)NH	0.3mM [U-99% 13C; U-99% 15N] PUB1 RRM3-16, 25mM potassium phosphate-17, 25mM sodium chloride-18, 0.5mM DTT-19, 0.1mM DSS-20	90% H2O/10% D2O		7.0	1 atm	298
8	3D CBCANH	0.3mM [U-99% 13C; U-99% 15N] PUB1 RRM3-16, 25mM potassium phosphate-17, 25mM sodium chloride-18, 0.5mM DTT-19, 0.1mM DSS-20	90% H2O/10% D2O		7.0	1 atm	298
9	3D HC -TOCSY	0.3mM [U-99% 13C; U-99% 15N] PUB1 RRM3-16, 25mM potassium phosphate-17, 25mM sodium chloride-18, 0.5mM DTT-19, 0.1mM DSS-20	90% H2O/10% D2O		7.0	1 atm	298
10	2D 1H-1H NOESY	0.3mM PUB1 RRM3-11, 25mM potassium phosphate-12, 25mM sodium chloride-13, 0.5mM DTT-14, 0.1mM DSS-15	100% D2O		7.0	1 atm	298
11	2D 1H-1H TOCSY	0.3mM PUB1 RRM3-6, 25mM potassium phosphate-7, 25mM sodium chloride-8, 0.5mM DTT-9, 0.01mM DSS-10	90% H2O/10% D2O		7.0	1 atm	298
12	2D 1H-1H TOCSY	0.3mM PUB1 RRM3-11, 25mM potassium phosphate-12, 25mM sodium chloride-13, 0.5mM DTT-14, 0.1mM DSS-15	100% D2O		7.0	1 atm	298
13	2D 1H-15N HSQC	0.3mM [U-99% 15N] PUB1 RRM3-1, 25mM potassium phosphate-2, 25mM sodium chloride-3, 0.5mM DTT-4, 0.01mM DSS-5	90% H2O/10% D2O		7.0	1 atm	298
14	2D 1H-13C HSQC	0.3mM [U-99% 13C; U-99% 15N] PUB1 RRM3-16, 25mM potassium phosphate-17, 25mM sodium chloride-18, 0.5mM DTT-19, 0.1mM DSS-20	90% H2O/10% D2O		7.0	1 atm	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	800

NMR Refinement
Method	Details	Software
torsion angle dynamics		TALOS

NMR Ensemble Information
Conformer Selection Criteria	target function
Conformers Calculated Total Number	50
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	TALOS		CORNILESCU, DELAGLIO
2	collection	TopSpin		Bruker Biospin
3	chemical shift assignment	CcpNmr Analysis		CCPN
4	peak picking	CcpNmr Analysis		CCPN
5	data analysis	CcpNmr Analysis		CCPN
6	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
7	structure solution	CYANA		Guntert, Mumenthaler and Wuthrich
8	structure analysis	MOLMOL		Koradi, Billeter and Wuthrich
9	refinement	CYANA

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.