2KQ2

Solution NMR structure of the apo form of a ribonuclease H domain of protein DSY1790 from Desulfitobacterium hafniense, Northeast Structural Genomics target DhR1A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.89 mM [U-99% 13C; U-99% 15N] DhR1A, 10 v/v [U-2H] D2O, 90 v/v H2O, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 50 uM DSS, 0.02 w/v sodium azide	90% H2O/10% D2O	430	6.5	ambient	298
2	2D 1H-13C HSQC	0.89 mM [U-99% 13C; U-99% 15N] DhR1A, 10 v/v [U-2H] D2O, 90 v/v H2O, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 50 uM DSS, 0.02 w/v sodium azide	90% H2O/10% D2O	430	6.5	ambient	298
3	2D 1H-13C HSQC	0.943 mM [U-5% 13C; U-99% 15N] DhR1A, 10 v/v [U-2H] D2O, 90 v/v H2O, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 50 uM DSS, 0.02 w/v sodium azide	90% H2O/10% D2O	430	6.5	ambient	298
4	3D CBCA(CO)NH	0.89 mM [U-99% 13C; U-99% 15N] DhR1A, 10 v/v [U-2H] D2O, 90 v/v H2O, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 50 uM DSS, 0.02 w/v sodium azide	90% H2O/10% D2O	430	6.5	ambient	298
5	3D HNCO	0.89 mM [U-99% 13C; U-99% 15N] DhR1A, 10 v/v [U-2H] D2O, 90 v/v H2O, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 50 uM DSS, 0.02 w/v sodium azide	90% H2O/10% D2O	430	6.5	ambient	298
6	3D HNCACB	0.89 mM [U-99% 13C; U-99% 15N] DhR1A, 10 v/v [U-2H] D2O, 90 v/v H2O, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 50 uM DSS, 0.02 w/v sodium azide	90% H2O/10% D2O	430	6.5	ambient	298
7	3D HBHA(CO)NH	0.89 mM [U-99% 13C; U-99% 15N] DhR1A, 10 v/v [U-2H] D2O, 90 v/v H2O, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 50 uM DSS, 0.02 w/v sodium azide	90% H2O/10% D2O	430	6.5	ambient	298
8	3D HCCH-COSY	0.89 mM [U-99% 13C; U-99% 15N] DhR1A, 10 v/v [U-2H] D2O, 90 v/v H2O, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 50 uM DSS, 0.02 w/v sodium azide	90% H2O/10% D2O	430	6.5	ambient	298
9	3D simultaneous NOESY	0.89 mM [U-99% 13C; U-99% 15N] DhR1A, 10 v/v [U-2H] D2O, 90 v/v H2O, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 50 uM DSS, 0.02 w/v sodium azide	90% H2O/10% D2O	430	6.5	ambient	298

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

NMR Refinement
Method	Details	Software
simulated annealing, torsion angle dynamics		VnmrJ

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

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VnmrJ		Varian
2	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	processing	NMRDraw		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	chemical shift assignment	XEASY		Bartels et al.
5	data analysis	XEASY		Bartels et al.
6	peak picking	XEASY		Bartels et al.
7	processing	SPSCAN		Glaser
8	chemical shift assignment	AutoAssign		Zimmerman, Moseley, Kulikowski and Montelione
9	structure solution	CSI		Wishart and Sykes
10	structure solution	TALOS		Cornilescu, Delaglio and Bax
11	structure solution	CYANA		Herrmann, Guntert and Wuthrich
12	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
13	data analysis	MOLMOL		Koradi, Billeter and Wuthrich
14	refinement	MOLMOL		Koradi, Billeter and Wuthrich
15	refinement	PSVS		Bhattacharya and 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.