1Q48

Solution NMR Structure of The Haemophilus Influenzae Iron-Sulfur Cluster Assembly Protein U (IscU) with Zinc Bound at the Active Site. Northeast Structural Genomics Consortium Target IR24. This protein is not apo, it is a model without zinc binding constraints.

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_15N-separated_NOESY	1 mM U-15N, U-13C IscU in 20 mM MES, 100 mM NaCl, 5 mM CaCl2, 10 mM DTT, 0.02% NaN3	95% H2O/5% D2O	20 mM MES, 100 mM NaCl, 5 mM CaCl2	6.5	ambient	293
2	3D_13C-separated_NOESY	1 mM U-15N, U-13C IscU in 20 mM MES, 100 mM NaCl, 5 mM CaCl2, 10 mM DTT, 0.02% NaN3	95% H2O/5% D2O	20 mM MES, 100 mM NaCl, 5 mM CaCl2	6.5	ambient	293
3	HNHA	1 mM U-15N, U-13C IscU in 20 mM MES, 100 mM NaCl, 5 mM CaCl2, 10 mM DTT, 0.02% NaN3	95% H2O/5% D2O	20 mM MES, 100 mM NaCl, 5 mM CaCl2	6.5	ambient	293
4	4D_13C-separated_NOESY	1 mM U-15N, U-13C IscU in 20 mM MES, 100 mM NaCl, 5 mM CaCl2, 10 mM DTT, 0.02% NaN3	100% D2O	20 mM MES, 100 mM NaCl, 5 mM CaCl2	6.5	ambient	293
5	13C_HSQC	1 mM U-15N, U-5%-13C IscU in 20 mM MES, 100 mM NaCl, 5 mM CaCl2, 10 mM DTT, 0.02% NaN3	95% H2O/5% D2O	20 mM MES, 100 mM NaCl, 5 mM CaCl2	6.5	ambient	293

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

NMR Refinement
Method	Details	Software
SIMULATED ANNEALING, TORSION ANGLE DYNAMICS, AUTOMATED ANALYSIS OF NOESY DATA AND 3D STRUCTURES	THE STRUCTURES ARE BASED ON A TOTAL OF 923 RESTRAINTS. SUMMARY OF EXPERIMENTAL CONSTRAINTS: DISTANCE CONSTRAINTS: TOTAL = 790; INTRA-RESIDUE [I=J] = 12; SEQUENTIAL [(I-J)=1] = 260; MEDIUM RANGE [1<(I-J)<5] = 185; LONG RANGE [(I-J)>=5] = 255; HYDROGEN BOND CONSTRAINTS = 58 (2 PER H-BOND); NUMBER OF DISTANCE CONSTRAINTS PER RESIDUE = 8.0; DIHEDRAL-ANGLE CONSTRAINTS = 133 (66 PHI, 67 PSI); TOTAL NUMBER OF CONSTRAINTS PER RESIDUE = 9.4 (RESIDES 26-123); NUMBER OF LONG RANGE CONSTRAINTS PER RESIDUE = 2.6; NUMBER OF STRUCTURES COMPUTED = 25; NUMBER OF STRUCTURES USED = 20. AVERAGE DISTANCE VIOLATIONS >0.0001 ANG = 24.3; AVERAGE R.M.S. DISTANCE VIOLATION = 0.003 ANG; MAXIMUM NUMBER OF DISTANCE VIOLATIONS 31. AVERAGE DIHEDRAL ANGLE VIOLATIONS: >0.0001 DEG = 1.0; MAX NUMBER OF DIHEDRAL ANGLE VIOLATIONS = 4; AVERAGE R.M.S. ANGLE VIOLATION = 0.01 DEG. RMSD VALUES: BACKBONE ATOMS (N,C,C',O) = 0.80 ANG; ALL HEAVY ATOMS = 1.17 ANG; PROCHECK: MOST FAVORED REGIONS = 77%; ADDITIONAL ALLOWED REGIONS = 20%; GENEROUSLY ALLOWED REGIONS = 3%; DISALLOWED REGIONS = 0%.	FELIX

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	25
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	THE STRUCTURE WAS DETERMINED USING TRIPLE-RESONANCE NMR SPECTROSCOPY, AUTOMATED ANALYSIS OF 3D STRUCTURE.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	FELIX	98	MSI (Accelrys)
2	structure solution	X-PLOR	Xplor-NIH-2.0.6	Schwieters, C.D., Kuszewski, J.J., Tjandra, N., Clore, G.M.
3	data analysis	Sparky	3.98	Goddard, T.D., Kneller, D.G.
4	refinement	XPLOR	XPLOR-NIH-2.0.6	SCHWIETERS, C.D., KUSZEWSKI, J.J. TJANDRA, N., CLORE, G.M.
5	refinement	AutoStructure		Huang, Y.J., Montelione G.T.
6	data analysis	TALOS	1999.019.15.47

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.