2AZH

Solution structure of iron-sulfur cluster assembly protein SUFU from Bacillus subtilis, with zinc bound at the active site. Northeast Structural Genomics Consortium target SR17

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 15N-1H HSQC, 2D 13C-1H HSQC, 3D HNCO, 3D HNCA, 3D HncoCA, 3D HNCACB, 3D HNcoCACB, 3D HcccoNH-TOCSY, 3D hCccoNH-TOCSY	0.7 MM U-15N, U-13C, PERDEUTERATEDSUFU WITH 13C,1H-LEU, VAL AND ILE-DELTA METHYLS, IN 50MM SODIUM PHOSPHATE, 100MM NACL, 1MM DTT, 0.02% NAN3	5% D2O, 95% H2O	100mM NaCl, 50mM Sodium Phosphate	6.5	ambient	20
2	2D HH NOESY, 2D 13C-1H HSQC, 2D 15N-1H HSQC, 3D 15N-NOESY, 3D 13C-NOESY, 4D CC NOESY	1.0 MM U-15N, U-13C, PERDEUTERATED SUFU WITH 13C,1H-LEU, VAL AND ILE-DELTA METHYLS AND 12C,15N,1H - PHE,TYR IN 100MM POTASSIUM PHOSPHATE, 200MM GLYCEROL, 1MM DTT, 0.02% NAN3	5% D2O, 95% H2O	100mM Potassium Phosphate	6.5	ambient	20
3	2D 13C-1H HSQC	1.0 MM U-5%13C, U-15N SUFU IN 100MM POTASSIUM PHOSPHATE, 200MM GLYCEROL, 1MM DTT, 0.02% NAN3	5% D2O, 95% H2O	100mM Potassium Phosphate	6.5	ambient	20
4	2D 15N-1H IPAP-HSQC	0.8 mM U-5%13C, U-15N SUFU IN 50MM SODIUM PHOSPHATE, 100MM NACL, 1MM DTT, 0.02% NAN3, 5% HEXAETHYLENE GLYCOL MONODODECYL ETHER/N-HEXANOL	90% H2O, 5% D2O	100mM NaCl, 50mM Sodium Phosphate	6.5	ambient	20
5	2D 15N-1H IPAP-HSQC	0.8 mM U-5%13C, U-15N SUFU IN 50MM SODIUM PHOSPHATE, 100MM NACL, 1MM DTT, 0.02% NAN3, 3.5% PENTAETHYLENE GLYCOL ETHER/N-OCTANOL	91.5% H2O, 5% D2O	100mM NaCl, 50mM Sodium Phosphate	6.5	ambient	20

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	500
2	Varian	INOVA	600
3	Varian	INOVA	750
4	Varian	INOVA	800

NMR Refinement
Method	Details	Software
MINIMAL CONSTRAINT STRUCTURE, CONTAINED 512 CONFORMATIONALLY RESTRICTING NOE-DERIVED DISTANCE CONSTRAINTS, 54 HYDROGEN BONDED PAIRS, 186 RESIDUAL DIPOLAR COUPLING CONSTRAINTS, 197 DIHEDRAL ANGLE CONSTRAINTS. 7.0 CONSTRAINTS PER CONFORMATIONALLY CONSTRAINED RESIDUE. 1.0 LONG RANGE CONSTRAINT PER RESIDUE.	DETERMINATION WAS PERFORMED WITH THE FOLLOWING STEPS: RESONANCE ASSIGNMENTS, TORTION ANGLE CONTRAINTS, HYDROGEN BONDED PAIRS AND NOESY CROSSPEAK DATA WAS USED AS INPUT INTO AUTOSTRUCTURE-DYANA. AUTOSTRUCTURE-DYANA IDENTIFIED DISTANCE CONSTRAINTS. THESE DISTANCE CONSTRAINTS WERE USED AS INPUT INTO AN XPLOR-NIH SIMULATED ANNEALING. USING CNS THE TOP TEN XPLOR-NIH STRUCTURES WERE ENERGY MINIMIZED IN EXPLICIT WATER WITH RDC CONSTRAINTS.	AUTOSTRUCTURE-DYANA

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)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	AUTOSTRUCTURE-DYANA	1.1.2	Huang, Montelione et al.
2	structure solution	XPLOR-NIH	2.0.6	Schwieters, Clore et al.
3	data analysis	TALOS	2.1	Cornilescu, Bax et al.
4	processing	NMRPipe	2.1	Delaglio, Bax et al.
5	data analysis	AutoAssign	1.9	Zimmerman, Montelione et al.
6	data analysis	Sparky	3.2	Goddard, Kneller et al.
7	data analysis	PdbStat	3.2	Tejero, Montelione et al.
8	data analysis	PALES	2.1	Zweckstetter
9	refinement	XPLOR-NIH	2.0.6

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.