1PP5

Structure of Antibacterial Peptide Microcin J25: a 21-Residue Lariat Protoknot

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_15N-separated_NOESY	2mM Microcin J25 U-13C,15N; 99.5% CD3OH, 0.5% H2O	99.5% CD3OH, 0.5% H2O	NA		ambient	298
2	3D_13C-separated_NOESY	2mM Microcin J25 U-13C,15N; 99.5% CD3OH, 0.5% H2O	99.5% CD3OH, 0.5% H2O	NA		ambient	298
3	HNHA	2mM Microcin J25 U-13C,15N; 99.5% CD3OH, 0.5% H2O	99.5% CD3OH, 0.5% H2O	NA		ambient	298
4	HNCACB	2mM Microcin J25 U-13C,15N; 99.5% CD3OH, 0.5% H2O	99.5% CD3OH, 0.5% H2O	NA		ambient	298
5	CBCAcoNH	2mM Microcin J25 U-13C,15N; 99.5% CD3OH, 0.5% H2O	99.5% CD3OH, 0.5% H2O	NA		ambient	298
6		2mM Microcin J25 U-13C,15N; 99.5% CD3OH, 0.5% H2O	99.5% CD3OH, 0.5% H2O	NA		ambient	298

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

NMR Refinement
Method	Details	Software
simulated annealing	The ensemble of structures is based on a total of 198 conformationally restraining constraints, 179 are NOE-derived distance constraints, 13 are dihedral angle constraints, 6 are distance constraints from hydrogen bonds.	AutoProc

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	60
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	The structure was determined using triple-resonance NMR spectroscopy.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	AutoProc	0.8	Bayro, M.J.; Monleon, D.; Baran, M.C.; Sahota, G.; Paranji, R.; Moseley, H.N.B.; Aramini, J.M.; Swapna, G.V.T.; Montelione, G.T.
2	processing	NMRPipe	1.4	Delaglio, F.; Grzesiek, S.; Vuister, G.; Zhu, G.; Pfeifer, J.; Bax, A.
3	structure solution	AutoStructure	1.1.2	Huang, Y.J., Tejero, R., Montelione, G.T.
4	structure solution	DYANA	1.5	Guntert, P.; Mumenthaler, C.; Wuthrich, K.
5	refinement	PDBStat	3.25	Tejero, R.; Montelione, G.T.

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.