2KA2

Solution NMR structure of BNIP3 transmembrane peptide dimer in detergent micelles with His173-Ser172 intermonomer hydrogen bond restraints

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D CN-NOESY-HSQC	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
2	3D 15N-edited NOESY-HSQC	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
3	3D 13C-edited NOESY-HSQC optimized for aromatic carbon	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
4	half-filtered 3D CN-NOESY-HSQC	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
5	2D spin-echo difference CT HSQC	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
6	HNHA	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
7	HNCA	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
8	CBCACONNH	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
9	HNCO	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
10	H(C)CH-COSY	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
11	(H)CCH-TOCSY	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
12	HBCBCGCDHD	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313
13	HBCBCGCDCEHE	0.4 mM [U-100% 13C; U-100% 15N] Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 0.6 mM Bcl2/Adenovirus E1B 19 kDa protein-interacting protein 3, 90% H2O, 10% D2O	90% H2O/10% D2O		5.1		313

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

NMR Refinement
Method	Details	Software
simulated annealing	Inferred intermonomer hydrogen bond restraints between His173-Ser172 are included in calculation. Intermonomer hydrogen bond restraints between His173-Ser172 are included. THE CALCULATION OF THIS STRUCTURE INCLUDES 2 ADDITIONAL HYDROGEN BOND RESTRAINTS THAT WERE DERIVED FROM MUTAGENESIS DATA.	ARIA

NMR Ensemble Information
Conformer Selection Criteria	lowest energy
Conformers Calculated Total Number	50
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	ARIA	1.2	Linge, O'Donoghue and Nilges
2	structure solution	ARIA	1.2	Linge, O'Donoghue and Nilges
3	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	data analysis	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
5	chemical shift assignment	Sparky		Goddard
6	peak picking	Sparky		Goddard
7	collection	VNMR		Varian

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.