2LLE

Computational design of an eight-stranded (beta/alpha)-barrel from fragments of different folds

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D HNCO	0.5 mM [U-100% 13C; U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
2	3D HNCA	0.5 mM [U-100% 13C; U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
3	3D HNCACB	0.5 mM [U-100% 13C; U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
4	3D C(CO)NH	0.5 mM [U-100% 13C; U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
5	3D CCH-TOCSY	0.5 mM [U-100% 13C; U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
6	3D CCH-COSY	0.5 mM [U-100% 13C; U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
7	2D PLUSH-TACSY	0.5 mM [U-100% 13C; U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
8	3D HNHA	0.5 mM [U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
9	3D HNHB	0.5 mM [U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
10	3D 1H-15N NOESY	0.5 mM [U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
11	3D 1H-13C NOESY	0.5 mM [U-100% 13C; U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
12	3D CNH-NOESY	0.5 mM [U-100% 13C; U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
13	3D CCH-NOESY	0.5 mM [U-100% 13C; U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
14	3D NNH-NOESY	0.5 mM [U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313
15	2D 15N-filtered 1H-1H NOESY	0.5 mM [U-100% 15N] CheYHisF-sfr_RM, 50 mM potassium phosphate, 300 mM potassium chloride	90% H2O/10% D2O	350	7.5	ambient	313

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

NMR Refinement
Method	Details	Software
simulated annealing	Non-bonded function including Ramachandran database potential	TopSpin

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	50
Conformers Submitted Total Number	17
Representative Model	1 (minimized average structure)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin		Bruker Biospin
2	processing	TopSpin		Bruker Biospin
3	chemical shift assignment	Sparky		Goddard
4	data analysis	Sparky		Goddard
5	structure solution	X-PLOR NIH	2.21	Schwieters, Kuszewski, Tjandra and Clore
6	refinement	X-PLOR NIH	2.21	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.