Experiment: 2NAE

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D 1H-15N NOESY	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
2	3D 1H-13C NOESY	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
3	3D 1H-15N NOESY (NOEs to lipid)	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
4	3D 1H-13C NOESY (methyl; double 13C-filter)	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
5	2D 1H-13C NOESY (aromatic; double 13C-filter)	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
6	2D 1H-15N HSQC	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
7	2D 1H-13C HSQC	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
8	3D HNCO	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
9	3D HNCA	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
10	3D HNCACB	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
11	3D CBCA(CO)NH	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
12	3D C(CO)NH	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
13	3D HBHA(CO)NH	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300
14	3D HCCH-TOCSY	0.5 mM [U-13C; U-15N] mCD28CD-1, 100 mM POPG-2, 125 mM DHPC-3, 20 mM sodium phosphate-4, 10 % [U-2H] D2O-5, 90 % H2O-6	90% H2O/10% D2O	20	6.7	ambient	300

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

NMR Refinement
Method	Details	Software
simulated annealing		TopSpin

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	200
Conformers Submitted Total Number	10
Representative Model	1 (fewest violations)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin	3.2	Bruker Biospin
2	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	data analysis	NMRView	5.2.2	Johnson, One Moon Scientific
4	data analysis	KUJIRA	0.9843	Naohiro Kobayashi
5	structure solution	X-PLOR NIH	2.32	Schwieters, Kuszewski, Tjandra and Clore
6	refinement	X-PLOR NIH	2.32	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.