2KEW

The solution structure of Bacillus subtilis SR211 START domain by NMR spectroscopy

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
2	2D 1H-13C HSQC	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
3	3D CBCA(CO)NH	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
4	3D C(CO)NH	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
5	3D HNCO	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
6	3D HNCA	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
7	3D HNCACB	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
8	3D HN(CO)CA	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
9	3D H(CCO)NH	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
10	3D HCCH-TOCSY	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
11	3D HNHA	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
12	3D HCCH-COSY	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
13	3D 1H-15N NOESY	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277
14	3D 1H-13C NOESY	1 mM [U-100% 13C; U-100% 15N] SR211, 20 mM MES, 5 % D2O, 0.02 % sodium azide, 10 mM DTT, 100 mM sodium chloride, 5 mM calcium chloride	95% H2O/5% D2O	0.105	6.5	ambient	277

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	Avance	600
2	Varian	INOVA	750
3	Varian	INOVA	600

NMR Refinement
Method	Details	Software
simulated annealing		ARIA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	70
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	The START domain is found in both eukaryotes and prokaryotes, with putative functions including signal transduction, transcriptional regulation, GTPase activation and thioester hydrolysis. Here we report the near complete 1H, 15N and 13C backbone and side chain NMR resonance assignments for the Bacillus subtilis START domain protein yndB.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	ARIA		Linge, O'Donoghue and Nilges
2	data analysis	ARIA		Linge, O'Donoghue and Nilges
3	refinement	ARIA		Linge, O'Donoghue and Nilges
4	chemical shift assignment	NMRView		Johnson, One Moon Scientific
5	data analysis	NMRView		Johnson, One Moon Scientific
6	peak picking	NMRView		Johnson, One Moon Scientific
7	processing	NMRDraw		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
8	data analysis	TALOS		Cornilescu, Delaglio and Bax

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.