Experiment: 2K9Y

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	3 mM [U-95% 15N] EphA2_TM 15N, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	90% H2O/10% D2O	10	5	AMBIENT	313
2	2D 1H-13C HSQC	3 mM [U-95% 13C; U-95% 15N] EphA2_TM 15N,13C, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	95% H2O/5% D2O	10	5	AMBIENT	313
3	3D HNCO	3 mM [U-95% 13C; U-95% 15N] EphA2_TM 15N,13C, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	95% H2O/5% D2O	10	5	AMBIENT	313
4	3D HNCA	3 mM [U-95% 13C; U-95% 15N] EphA2_TM 15N,13C, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	95% H2O/5% D2O	10	5	AMBIENT	313
5	3D HCCH-TOCSY	3 mM [U-95% 13C; U-95% 15N] EphA2_TM 15N,13C, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	95% H2O/5% D2O	10	5	AMBIENT	313
6	3D 1H-15N NOESY	3 mM [U-95% 15N] EphA2_TM 15N, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	90% H2O/10% D2O	10	5	AMBIENT	313
7	3D 1H-15N TOCSY	3 mM [U-95% 15N] EphA2_TM 15N, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	90% H2O/10% D2O	10	5	AMBIENT	313
8	3D 1H-13C NOESY	3 mM [U-95% 13C; U-95% 15N] EphA2_TM 15N,13C, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	95% H2O/5% D2O	10	5	AMBIENT	313
9	3D HNHB	3 mM [U-95% 15N] EphA2_TM 15N, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	90% H2O/10% D2O	10	5	AMBIENT	313
10	3D HNHA	3 mM [U-95% 15N] EphA2_TM 15N, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	90% H2O/10% D2O	10	5	AMBIENT	313
11	2D 1H-1H NOESY	3 mM [U-95% 15N] EphA2_TM 15N, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	90% H2O/10% D2O	10	5	AMBIENT	313
12	13C F1-filtered/F3-edited-NOESY	1.5 mM [U-95% 13C; U-95% 15N] EphA2_TM 15N,13C, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer, 1.5 mM EphA2_TM	100% D2O	10	5	AMBIENT	313
13	15N-T2	3 mM [U-95% 15N] EphA2_TM 15N, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	90% H2O/10% D2O	10	5	AMBIENT	313
14	15N-T1	3 mM [U-95% 15N] EphA2_TM 15N, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	90% H2O/10% D2O	10	5	AMBIENT	313
15	15N-NOE	3 mM [U-95% 15N] EphA2_TM 15N, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	90% H2O/10% D2O	10	5	AMBIENT	313
16	2D 1H-1H TOCSY	3 mM [U-95% 15N] EphA2_TM 15N, 96 mM [U-2H] DHPC, 24 mM [U-2H] DMPC, 1.5 mM NaN3, 1 mM EDTA, 10 mM phosphate buffer	90% H2O/10% D2O	10	5	AMBIENT	313

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	UNITY	600
2	Varian	INOVA	600

NMR Refinement
Method	Details	Software
TORSION ANGLE DYNAMICS, molecular dynamics, TORSION ANGLE DYNAMICS	First 15 models represents Torsion Angle Dynamics with applied NMR restrains using CYANA software, Last two models (16 and 17) represents MD relaxation in the explicit hydrated DPPC bilayer with NMR restrains using GROMACS software	NMRPipe

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	100
Conformers Submitted Total Number	17
Representative Model	1 (fewest violations)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	spectra processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	assignment	CARA	1.5.5, 1.8	Keller and Wuthrich
3	structure calculation	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
4	collection	VNMR		Varian
5	geometry optimization	GROMACS	3.3.2	Erik Lindahl
6	refinement	GROMACS	3.3.2	Erik Lindahl

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.