1ZU2

Solution NMR structure of the plant Tom20 mitochondrial import receptor from Arabidopsis thaliana

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_13C-separated_NOESY	1mM Tom20 U-15N,13C, 20mM sodium phosphate buffer, 150mM sodium chloride, 0.02 %(w/v) sodium azide, 1mM dithiothreitol, 1mM EDTA, 3mM phenylmethylsulphonylfluoride, 90 % H2O, 10 % D2O	90% H2O/10% D2O	150mM sodium chloride	7.4	ambient	298
2	3D_15N-separated_NOESY	1mM Tom20 U-15N, 20mM sodium phosphate buffer, 150mM sodium chloride, 0.02 %(w/v) sodium azide, 1mM dithiothreitol, 1mM EDTA, 3mM phenylmethylsulphonylfluoride, 90 % H2O, 10 % D2O	90% H2O/10% D2O	150mM sodium chloride	7.4	ambient	298
3	HNHA	1mM Tom20 U-15N, 20mM sodium phosphate buffer, 150mM sodium chloride, 0.02 %(w/v) sodium azide, 1mM dithiothreitol, 1mM EDTA, 3mM phenylmethylsulphonylfluoride, 90 % H2O, 10 % D2O	90% H2O/10% D2O	150mM sodium chloride	7.4	ambient	298
4	3D-13C-separated_NOESY (aromatics)	1mM Tom20 U-15N,13C, 20mM sodium phosphate buffer, 150mM sodium chloride, 0.02 %(w/v) sodium azide, 1mM dithiothreitol, 1mM EDTA, 3mM phenylmethylsulphonylfluoride	95 % D2O	150mM sodium chloride	7.4	ambient	298
5	3D_13C-separated_NOESY	1mM Tom20 U-15N,13C, 20mM sodium phosphate buffer, 150mM sodium chloride, 0.02 %(w/v) sodium azide, 1mM dithiothreitol, 1mM EDTA, 3mM phenylmethylsulphonylfluoride	95 % D2O	150mM sodium chloride	7.4	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600
2	Varian	INOVA	500

NMR Refinement
Method	Details	Software
torsion angle dynamics	The CANDID software was used for initial automatic NOE assignment. Structures are based on 1629 NOE-derived upper distance constraints (47 intra residue), 136 distance restraints from hydrogen bonds, refinement against 129 J(HNHA) coupling constants and 234 PHI/PSI dihedral angle constraints from TALOS.	VNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	Structure contains 5 non-native residues at the N-terminus (from fusion system protease site) and 8 non-native residues at the C-terminus from a polyhistidine tag.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VNMR	4.1c	Varian Inc.
2	processing	NMRPipe	1.1	Delaglio, F.
3	data analysis	Sparky	3.111	Goddard, T.D & Kneller, D.G.
4	structure solution	CYANA	1.0.7	Guentert, P.
5	data analysis	CANDID	1.0	Guentert, P. & Herrmann, T.
6	refinement	CYANA	1.0.7	Guentert, P.

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.