1R57

NMR Solution Structure of a GCN5-like putative N-acetyltransferase from Staphylococcus aureus. Northeast Structural Genomics Consortium Target ZR31

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_13C-separated_NOESY	1 mM protein U-15N,13C; 20 mM MES, 100 mM NaCl, 5 mM CaCl2, 0.02% NaN3; 95% H2O, 5% D2O	95% H2O/5% D2O	.115	6.5	ambient	293
2	3D_15N-separated_NOESY	1 mM protein U-15N,13C; 20 mM MES, 100 mM NaCl, 5 mM CaCl2, 0.02% NaN3; 95% H2O, 5% D2O	95% H2O/5% D2O	.115	6.5	ambient	293
3	HNHA	1 mM protein U-15N,13C; 20 mM MES, 100 mM NaCl, 5 mM CaCl2, 0.02% NaN3; 95% H2O, 5% D2O	95% H2O/5% D2O	.115	6.5	ambient	293
4	HNHB	0.4 mM protein U-15N,5%-13C; 20 mM MES, 100 mM NaCl, 5 mM CaCl2, 0.02% NaN3; 95% H2O, 5% D2O	95% H2O/5% D2O	.115	6.5	ambient	293
5	4D_13C-separated_NOESY	1 mM protein U-15N,13C, 20 mM MES, 100 mM NaCl, 5 mM CaCl2, 0.02% NaN3; 100% D2O	100% D2O	.115	6.5	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600
2	Varian	INOVA	750

NMR Refinement
Method	Details	Software
automated generation of initial distance restraint set distance geometry simulated annealing final refinement in explicit solvent	Backbone and sidechain assignments were determined manually from triple-resonance NMR data. NOE distance restraints were derived automatically from peak-picked data with AutoStructure, then error-checked and corrected manually. The structure is based on 848 restraints: 710 meaningful distance restraints, 58 hydrogen bond restraints, and 80 dihedral angle restraints. There are 9.5 restraints per restrained residue. Phi dihedral restraints were derived from the HNHA experiment and TALOS. Psi dihedral restraints were derived from NOE ratios, secondary structure propensities evident in preliminary structures, alpha carbon chemical shifts, and TALOS. Residues 44-51 comprise a poorly-defined loop in this ensemble of structures. Residues 1-3 and 94-102 are unstructured termini.	AutoStructure

NMR Ensemble Information
Conformer Selection Criteria	structures with fewest restraint violations and lowest energy
Conformers Calculated Total Number	30
Conformers Submitted Total Number	20
Representative Model	1 (similarity to average, few violations, and low energy)

Additional NMR Experimental Information
Details	amide proton exchange was measured by dissolving a lyophilized protonated sample in D2O

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	data analysis	AutoStructure	1.1	Huang, Tejero, Montelione
2	structure solution	CNS	1.1	Brunger, Schwieters, Kuszewski, Tjandra, Clore
3	refinement	CNS	1.1	Brunger
4	processing	Felix	97	MSI
5	data analysis	Sparky	3	Goddard, Kneller
6	data analysis	TALOS		Cornilescu, Delaglio, Bax

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.