Experiment: 2ROP

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.1-0.4mM [U-99% 13C; U-99% 15N] third soluble domain; 0.1-0.4mM [U-99% 13C; U-99% 15N] fourth soluble domain; 90% H2O/10% D2O	90% H2O/10% D2O	20mM Na Pi + 150mM NaCl	6.5	ambient	298
2	3D CBCA(CO)NH	0.1-0.4mM [U-99% 13C; U-99% 15N] third soluble domain; 0.1-0.4mM [U-99% 13C; U-99% 15N] fourth soluble domain; 90% H2O/10% D2O	90% H2O/10% D2O	20mM Na Pi + 150mM NaCl	6.5	ambient	298
3	3D HNCACB	0.1-0.4mM [U-99% 13C; U-99% 15N] third soluble domain; 0.1-0.4mM [U-99% 13C; U-99% 15N] fourth soluble domain; 90% H2O/10% D2O	90% H2O/10% D2O	20mM Na Pi + 150mM NaCl	6.5	ambient	298
4	3D HCCH-TOCSY	0.1-0.4mM [U-99% 13C; U-99% 15N] third soluble domain; 0.1-0.4mM [U-99% 13C; U-99% 15N] fourth soluble domain; 90% H2O/10% D2O	90% H2O/10% D2O	20mM Na Pi + 150mM NaCl	6.5	ambient	298
5	3D 1H-15N NOESY	0.1-0.4mM [U-99% 13C; U-99% 15N] third soluble domain; 0.1-0.4mM [U-99% 13C; U-99% 15N] fourth soluble domain; 90% H2O/10% D2O	90% H2O/10% D2O	20mM Na Pi + 150mM NaCl	6.5	ambient	298
6	3D 1H-13C NOESY	0.1-0.4mM [U-99% 13C; U-99% 15N] third soluble domain; 0.1-0.4mM [U-99% 13C; U-99% 15N] fourth soluble domain; 90% H2O/10% D2O	90% H2O/10% D2O	20mM Na Pi + 150mM NaCl	6.5	ambient	298
7	R1	0.1-0.4mM [U-99% 13C; U-99% 15N] third soluble domain; 0.1-0.4mM [U-99% 13C; U-99% 15N] fourth soluble domain; 90% H2O/10% D2O	90% H2O/10% D2O	20mM Na Pi + 150mM NaCl	6.5	ambient	298
8	R2	0.1-0.4mM [U-99% 13C; U-99% 15N] third soluble domain; 0.1-0.4mM [U-99% 13C; U-99% 15N] fourth soluble domain; 90% H2O/10% D2O	90% H2O/10% D2O	20mM Na Pi + 150mM NaCl	6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	900
2	Bruker	AVANCE	700
3	Bruker	AVANCE	500

NMR Refinement
Method	Details	Software
simulated annealing	The construct investigated consists of domains 3 and 4 of the soluble N-terminal tail of ATP7B and of a inter-domain linker of 31 aa being essentially in a random coil form. The two domains reorient in solution independently of one another and the coordinates of the linker have been omitted.	CYANA

NMR Ensemble Information
Conformer Selection Criteria	target function
Conformers Calculated Total Number	400
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	data analysis	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
2	chemical shift assignment	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
3	data analysis	Amber	8	Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... and Kollm
4	chemical shift assignment	Amber	8	Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... and Kollm
5	data analysis	CYANA	2.1	Keller and Wuthrich
6	chemical shift assignment	CYANA	2.1	Keller and Wuthrich
7	refinement	CYANA	2.1	Guntert, Mumenthaler and Wuthrich

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.