2KVS

NMR Solution Structure of Q7A1E8 protein from Staphylococcus aureus: Northeast Structural Genomics Consortium target: ZR215

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293
2	2D 1H-13C HSQC	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293
3	3D CBCA(CO)NH	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293
4	3D HNCACB	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293
5	3D HBHA(CO)NH	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293
6	3D HNCO	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293
7	3D HNHA	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293
8	3D HCCH-TOCSY	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293
9	3D 1H-15N NOESY	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293
10	3D 1H-13C NOESY	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293
11	2D 1H-15N HNOE	0.71 mM [U-100% 13C; U-100% 15N] ZR215-1	90% H2O/10% D2O	100mM NaCl	7.5	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600
2	Bruker	AVANCE	800

NMR Refinement
Method	Details	Software
simulated annealing	400 structures were calculated and 20 best conformers were then refined in a shell of water using CNS. Initial dihedral angle constriants were obtained from TALOS. Final quality factors determined using PSVS software. Ordered residues were defined as 8-12,18-30,35-37,43-77.RMSD(ordered residues):Backbone 1.0A, All atoms 1.5A; Ramachandran statistics for all ordered regions: Most favored regions: 91.5%; Additionally allowed: 8.4% and generously allowed: 0.1%. Procheck scores for all ordered residues are (Raw/Z): phi-psi 0.11/0.75 All:0.04/0.24; MolProbity clash score(Raw/Z):13.45/-0.78; RPF scores for goodness of fit of NOESY data: Recall: 0.954; Precision:0.928; F-measure: 0.941; DP-score 0.798.	AutoAssign

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	400
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 resonance assignments were made using AutoAssign. Sidechain assignments were completed manually. Chemical shift assignments were validated using AVS software. Automated NOESY assignments were made using AutoStructure and structure solution was determined using CYANA-2.1.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	chemical shift assignment	AutoAssign	2.1	Zimmerman, Moseley, Kulikowski and Montelione
2	processing	NMRPipe	2008	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	peak picking	Sparky	2.3	Goddard
4	data analysis	Sparky	2.3	Goddard
5	collection	VnmrJ	2.1B	Varian
6	collection	TopSpin	2.1	Bruker Biospin
7	structure solution	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
8	data analysis	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
9	geometry optimization	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
10	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
11	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
12	chemical shift validation	AVS		Moseley and Montelione
13	structure validation	PSVS	1.3	Bhattacharya and Montelione
14	dihedral angle constriants	TALOS		Cornilescu, Delaglio and Bax

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.