2N2T

Solution NMR Structure of DE NOVO DESIGNED PROTEIN (FDA_60), Northeast Structural Genomics Consortium (NESG) Target OR303

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.87 mM [U-100% 13C; U-100% 15N] OR303.005	90% H2O/10% D2O		4.5	ambient	298
2	2D 1H-13C HSQC	0.87 mM [U-100% 13C; U-100% 15N] OR303.005	90% H2O/10% D2O		4.5	ambient	298
3	3D HNCO	0.87 mM [U-100% 13C; U-100% 15N] OR303.005	90% H2O/10% D2O		4.5	ambient	298
4	3D CBCA(CO)NH	0.87 mM [U-100% 13C; U-100% 15N] OR303.005	90% H2O/10% D2O		4.5	ambient	298
5	3D HNCACB	0.87 mM [U-100% 13C; U-100% 15N] OR303.005	90% H2O/10% D2O		4.5	ambient	298
6	3D 1H-13C arom NOESY	0.87 mM [U-100% 13C; U-100% 15N] OR303.005	90% H2O/10% D2O		4.5	ambient	298
7	3D simutaneous 13C-aromatic,13C-aliphatic,15N edited 1H-1H NOESY	0.87 mM [U-100% 13C; U-100% 15N] OR303.005	90% H2O/10% D2O		4.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	800
2	Varian	INOVA	600

NMR Refinement
Method	Details	Software
distance geometry, simulated annealing, molecular dynamics		CNS

NMR Ensemble Information
Conformer Selection Criteria	target function
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	geometry optimization	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
3	refinement	AutoStructure	2.1	Huang, Tejero, Powers and Montelione
4	chemical shift assignment	AutoAssign	2.1	Zimmerman, Moseley, Kulikowski and Montelione
5	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
6	peak picking	XEASY		Bartels et al.
7	collection	TopSpin		Bruker Biospin
8	collection	VnmrJ		Varian
9	data analysis	Sparky		Goddard
10	geometry optimization	TALOS+		Shen, Cornilescu, Delaglio and Bax
11	data analysis	PSVS		Bhattacharya, Montelione
12	refinement	CYANA		Guntert, Mumenthaler and Wuthrich
13	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read

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.