2MOQ

Solution Structure and Molecular determinants of Hemoglobin Binding of the first NEAT Domain of IsdB in Staphylococcus aureus

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1 mM [U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 5 % [U-99% 2H] D2O, 95 % H2O	95% H2O/5% D2O	0.45	6.8	ambient	298
2	3D 1H-15N TOCSY	1 mM [U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 5 % [U-99% 2H] D2O, 95 % H2O	95% H2O/5% D2O	0.45	6.8	ambient	298
3	3D 1H-15N NOESY	1 mM [U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 5 % [U-99% 2H] D2O, 95 % H2O	95% H2O/5% D2O	0.45	6.8	ambient	298
4	3D CBCA(CO)NH	1 mM [U-99% 13C; U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 5 % [U-99% 2H] D2O, 95 % H2O	95% H2O/5% D2O	0.45	6.8	ambient	298
5	3D C(CO)NH	1 mM [U-99% 13C; U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 5 % [U-99% 2H] D2O, 95 % H2O	95% H2O/5% D2O	0.45	6.8	ambient	298
6	3D HNCA	1 mM [U-99% 13C; U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 5 % [U-99% 2H] D2O, 95 % H2O	95% H2O/5% D2O	0.45	6.8	ambient	298
7	3D HNCACB	1 mM [U-99% 13C; U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 5 % [U-99% 2H] D2O, 95 % H2O	95% H2O/5% D2O	0.45	6.8	ambient	298
8	3D HBHA(CO)NH	1 mM [U-99% 13C; U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 5 % [U-99% 2H] D2O, 95 % H2O	95% H2O/5% D2O	0.45	6.8	ambient	298
9	3D H(CCO)NH	1 mM [U-99% 13C; U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 5 % [U-99% 2H] D2O, 95 % H2O	95% H2O/5% D2O	0.45	6.8	ambient	298
10	3D HCCH-TOCSY	1 mM [U-99% 13C; U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 100 % [U-99% 2H] D2O	100% D2O	0.45	6.8	ambient	298
11	3D 1H-13C NOESY	1 mM [U-99% 13C; U-99% 15N] IsdB N1, 50 mM sodium phosphate, 400 mM sodium chloride, 0.1 mM PMSF, 1 mM EDTA, 0.01 % sodium azide, 100 % [U-99% 2H] D2O	100% D2O	0.45	6.8	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	600

NMR Refinement
Method	Details	Software
torsion angle dynamics, molecular dynamics		TopSpin

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	200
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin	2.1	Bruker Biospin
2	processing	NMRPipe	7.4	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	processing	NMRDraw	7.4	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	data analysis	Sparky	3.113	Goddard
5	peak picking	Sparky	3.113	Goddard
6	chemical shift assignment	Sparky	3.113	Goddard
7	refinement	CYANA		Guntert, Mumenthaler and Wuthrich
8	refinement	Amber		Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... and Kollman

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.