2LNU

Solution NMR Structure of the uncharacterized protein from gene locus rrnAC0354 of Haloarcula marismortui, Northeast Structural Genomics Consortium Target HmR11

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.7 mM [U-100% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
2	2D 1H-13C HSQC	0.7 mM [U-100% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
3	3D HNCO	0.7 mM [U-100% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
4	3D CBCA(CO)NH	0.7 mM [U-100% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
5	3D HNCACB	0.7 mM [U-100% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
6	3D 1H-13C arom NOESY	0.7 mM [U-100% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
7	3D 13C-aromatic,13C-aliphatic,15N edited 1H-1H NOESY	0.7 mM [U-100% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
8	2D hetnoe	1.2 mM [U-5% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
9	1D T1 inv.rec	1.2 mM [U-5% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
10	1D T2 CPMG	1.2 mM [U-5% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
11	2D 1H-13C HSQC highres	1.2 mM [U-5% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
12	Jmod 15N TROSY	0.75 mM [U-5% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298
13	jmod 15N TROSY	0.75 mM [U-5% 13C; U-100% 15N] HmR11, 0.02 % NaN3, 10 mM DTT, 5 mM CaCL2, 100 mM NaCL, 1 x Proteinase Inhibitors, 20 mM MES pH 6.5, 10 % D2O, 50 uM DSS	95% H2O/5% D2O		6.5	ambient	298

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

NMR Refinement
Method	Details	Software
molecular dynamics	cns with rdc noe and dihedral constraints	CNS

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)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
2	structure solution	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
3	geometry optimization	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
4	refinement	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
5	geometry optimization	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
6	structure solution	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
7	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
8	data analysis	XEASY		Bartels et al.
9	peak picking	XEASY		Bartels et al.
10	chemical shift assignment	XEASY		Bartels et al.
11	collection	TOPSPIN		Bruker Biospin
12	collection	VNMRJ		Varian
13	chemical shift assignment	PINE		Bahrami, Markley, Assadi, and Eghbalnia
14	data analysis	SPARKY		Goddard
15	geometry optimization	TALOS+		Shen, Cornilescu, Delaglio and Bax
16	geometry optimization	PALES		PALES (Zweckstetter, Bax)
17	structure validation	PSVS		Bhattacharya, Montelione

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.