2K4V

Solution structure of uncharacterized protein PA1076 from Pseudomonas aeruginosa. Northeast Structural Genomics Consortium (NESG) target PaT3, Ontario Center for Structural Proteomics target PA1076 .

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
2	2D 1H-13C HSQC	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
3	3D CBCA(CO)NH	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
4	3D HBHA(CO)NH	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
5	3D HNCO	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
6	3D HNCA	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
7	3D C(CO)NH	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
8	3D H(CCO)NH	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
9	3D HCCH-TOCSY	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
10	3D HCCH-TOCSY	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
11	3D 1H-15N NOESY	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
12	3D 1H-13C NOESY	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
13	3D 1H-13C aromatic NOESY	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
14	3D H(C)CH-TOCSY aromatic	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
15	3D (H)CCH-TOCSY aromatic	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
16	T1rho array	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
17	3D IPAP E-COSY HNCO	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298
18	3D IPAP E-COSY HNCO	0.7-1 mM [U-100% 13C; U-100% 15N] PA1076, 1x inhibitor cocktail (purchased from Roche), 10 mM [U-2H] TRIS, 300 mM sodium chloride, 0.01 % sodium azide, 1 mM benzamidine	90% H2O/10% D2O	300	7	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	800
2	Bruker	AVANCE	600
3	Bruker	AVANCE	500

NMR Refinement
Method	Details	Software
simulated annealing	water refinement with RDC constraints	CcpNmr Analysis

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	5 mm Shigemi tube Most 3D spectra collected using non-uniform sampling and processed with MDDNMR

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	peak picking	CcpNmr Analysis	1.4	CCPN
2	chemical shift assignment	CcpNmr Analysis	1.4	CCPN
3	processing	MddNMR	1.2c	V. Orekhov, I. Ibraghimov
4	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
5	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
6	geometry optimization	TALOS		Cornilescu, Delaglio and Bax
7	chemical shift assignment	ABACUS		A. Lemak
8	noe assignment	ABACUS		A. Lemak
9	refinement	CNSSOLVE		Brunger, Adams, Clore, Gros, Nilges and Read

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.