Experiment: 5X29

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.67 mM [U-99% 15N] Envelope small membrane protein, 200 mM LMPG, 20 mM sodium phosphate, 50 mM sodium chloride	95% H2O/5% D2O	50 mM	5.5	1 atm	308	Bruker AVANCE II 700
2	3D HNCA	0.67 mM [U-99% 13C; U-99% 15N] Envelope small membrane protein, 200 mM LMPG, 20 mM sodium phosphate, 50 mM sodium chloride	95% H2O/5% D2O	50 mM	5.5	1 atm	308	Bruker AVANCE II 700
3	3D HN(CO)CA	0.67 mM [U-99% 13C; U-99% 15N] Envelope small membrane protein, 200 mM LMPG, 20 mM sodium phosphate, 50 mM sodium chloride	95% H2O/5% D2O	50 mM	5.5	1 atm	308	Bruker AVANCE II 700
4	3D HCCH-TOCSY	0.67 mM [U-99% 13C; U-99% 15N] Envelope small membrane protein, 200 mM LMPG, 20 mM sodium phosphate, 50 mM sodium chloride	95% H2O/5% D2O	50 mM	5.5	1 atm	308	Bruker AVANCE II 700
5	3D 1H-15N NOESY	0.67 mM [U-99% 13C; U-99% 15N] Envelope small membrane protein, 200 mM LMPG, 20 mM sodium phosphate, 50 mM sodium chloride	95% H2O/5% D2O	50 mM	5.5	1 atm	308	Bruker AVANCE II 700
7	2D 1H-13C HSQC	0.67 mM [U-99% 13C; U-99% 15N] Envelope small membrane protein, 200 mM LMPG, 20 mM sodium phosphate, 50 mM sodium chloride	95% H2O/5% D2O	50 mM	5.5	1 atm	308	Bruker AVANCE II 700
6	3D 1H-13C NOESY	0.67 mM [U-99% 13C; U-99% 15N] Envelope small membrane protein, 200 mM LMPG, 20 mM sodium phosphate, 50 mM sodium chloride	95% H2O/5% D2O	50 mM	5.5	1 atm	308	Bruker AVANCE II 700
8	3D 1H-15N NOESY	0.67 mM [U-99% 15N, U-99% 2H] Envelope small membrane protein, 200 mM LMPG, 20 mM sodium phosphate, 50 mM sodium chloride	95% H2O/5% D2O	50 mM	5.5	1 atm	308	Bruker AVANCE II 700
9	3D 1H-15N NOESY	0.33 mM [U-99% 13C] Envelope small membrane protein, 0.33 mM [U-99% 15N, U-99% 2H] Envelope small membrane protein, 200 mM LMPG, 20 mM sodium phosphate, 50 mM sodium chloride	95% H2O/5% D2O	50 mM	5.5	1 atm	308	Bruker AVANCE II 700

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE II	700

NMR Refinement
Method	Details	Software
simulated annealing	Calculation of pentamer structure	HADDOCK

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	16
Representative Model	1 (medoid)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin		Bruker Biospin
2	chemical shift assignment	CARA		Keller and Wuthrich
3	structure calculation	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
4	refinement	HADDOCK	2.2	Bonvin

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 R01GM157729.