2KKO

Solution NMR structure of the homodimeric winged helix-turn-helix DNA-binding domain (fragment 1-100) Mb0332 from Mycobacterium bovis, a possible ArsR-family transcriptional regulator. Northeast Structural Genomics Consortium Target MbR242E.

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
2	2D 1H-13C HSQC	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
3	3D 1H-15N NOESY	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
4	3D 1H-13C NOESY	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
5	3D HNCO	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
6	3D HNCA	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
7	3D CBCA(CO)NH	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
8	3D HNCACB	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
9	3D C(CO)NH	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
10	3D HBHA(CO)NH	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
11	3D HCCH-TOCSY	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
12	3D HCCH-COSY	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
13	2D 1H-13C HSQC	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 0.8 mM [U-5% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
14	3D 1H-13C NOESY arom	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 1.1 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS	95% H2O/5% D2O	200	6.0	ambient	293
15	3D CN filt 1H-13C NOESY	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 0.5 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS, .5 mM protein	95% H2O/5% D2O	200	6.0	ambient	293
16	2D 1H-13C HSQC	20 mM MES, 200 mM sodium chloride, 10 mM DTT, 0.02 % sodium azide, 0.5 mM [U-100% 13C; U-100% 15N] protein, 50 uM DSS, .5 mM protein	95% H2O/5% D2O	200	6.0	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600
2	Bruker	AVANCE III	850
3	Varian	INOVA	750

NMR Refinement
Method	Details	Software
simulated annealing	CNS water refinement	NMRPipe

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	150
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	NMRPipe	2008	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	collection	VNMR	6.1C	Varian
3	collection	TopSpin	2.1.3	Bruker Biospin
4	data analysis	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
5	structure solution	X-PLOR NIH	2.20	Schwieters, Kuszewski, Tjandra and Clore
6	data analysis	Sparky	3.113	Goddard
7	structure validation	PSVS	1.3	Bhattacharya and Montelione
8	chemical shift assignment	AutoAssign	2.3.0	Zimmerman, Moseley, Kulikowski and Montelione
9	data analysis	PdbStat	5.1	(PDBStat) R. Tejero, G.T. Montelione
10	refinement	CNS	1.2	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.