Experiment: 1Z2Q

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_13C-separated_NOESY	0.8mM LM5-1 U-15N, 13C; 50mM sodium phosphate buffer, 50mM KCl, 1mM DTT, 1mM AEBSF, 0.05% sodium azide; 90% H2O, 10% D2O	90% H2O/10% D2O		7.4	1 atm	293
2	3D_15N-separated_NOESY	0.8mM LM5-1 U-15N, 13C; 50mM sodium phosphate buffer, 50mM KCl, 1mM DTT, 1mM AEBSF, 0.05% sodium azide; 90% H2O, 10% D2O	90% H2O/10% D2O		7.4	1 atm	293
3	HNHA	0.8mM LM5-1 U-15N, 13C; 50mM sodium phosphate buffer, 50mM KCl, 1mM DTT, 1mM AEBSF, 0.05% sodium azide; 90% H2O, 10% D2O	90% H2O/10% D2O		7.4	1 atm	293
4	3D_13C-separated_NOESY	0.8mM LM5-1 U-15N,13C; 50mM sodium phosphate buffer, 50mM KCl, 1mM DTT, 1mM AEBSF,0.05% sodium azide; 100% D2O	99.99% D2O		7.4	1 atm	293
5	2D_13C-ct-HSQC	0.8mM LM5-1 ~10% 13C; 50mM sodium phosphate buffer, 50mM KCl, 1mM DTT, 1mM AEBSF, 0.05% sodium azide; 100% D2O	90% H2O/10% D2O		7.4	1 atm	293
6	2D_15N-HSQC-TROSY, 2D_HN(a/b-NC-J)-TROSY, 2D_HN(CO-a/b-C'CA-J)-TROSY, 3D_1H-dec_HN(CO)CA	0.8mM LM5-1 U-15N,13C; 50mM sodium phosphate buffer, 50mM KCl, 1mM DTT, 1mM AEBSF, 0.05% sodium azide; 10mg/ml PF1; 90% H2O, 10% D2O	90% H2O/10% D2O		7.4	1 atm	293

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

NMR Refinement
Method	Details	Software
torsion angle dynamics, simulated annealing.	Structures based on 545 NOE-derived distance constraints, 101 dihedral angle restraints, and 259 residual dipolar couplings.	VNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations, structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	21
Representative Model	21 (minimized average structure)

Additional NMR Experimental Information
Details	2 x Zn2+ ions not included in calculations, ligating cysteines treated as protonated sulfhydryl groups during simulated annealing.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VNMR	6.1c	Varian inc.
2	processing	NMRPipe	2.2	Delaglio
3	data analysis	XEASY	1.3	Bartels
4	structure solution	CYANA	1.0.7	Guntert
5	refinement	Xplor-NIH	2.9.1

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.