2LEK

Solution NMR structure of a Thiamine Biosynthesis (ThiS) Protein RPA3574 from Rhodopseudomonas palustris refined with NH RDCs. Northeast Structural Genomics Consortium target RpR325

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
2	2D 1H-13C HSQC aliphatic	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
3	2D 1H-13C HSQC aromatic	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
4	2D 1H-13C HSQC-CT	0.8 mM U-100% 15N and 5% 13C biosynthetically directed protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide	90% H2O/10% D2O	0.1	6.5	ambient	293
5	3D 1H-15N NOESY	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
6	3D 1H-13C NOESY_aliph	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
7	3D HNCO	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
8	3D HNCACB	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
9	3D CBCA(CO)NH	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
10	3D 1H-13C NOESY_arom	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
11	3D HBHA(CO)NH	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
12	3D C(CCO)NH	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
13	3D HCCH-TOCSY	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
14	3D CCH-TOCSY	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	100% D2O	0.1	6.5	ambient	293
15	2D 1H-15N HSQC_His	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
16	3D HNCA	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293
17	3D HN(CO)CA	0.9 mM [U-100% 13C; U-100% 15N] protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.02 % sodium azide, 10 mM DTT	90% H2O/10% D2O	0.1	6.5	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600
2	Bruker	AVANCE III	850
3	Varian	INOVA	500

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	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

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	2.1.4	Bruker Biospin
4	data analysis	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
5	structure solution	X-PLOR NIH	2.25	Schwieters, Kuszewski, Tjandra and Clore
6	refinement	CNS	1.3	Brunger, Adams, Clore, Gros, Nilges and Read
7	data analysis	Sparky	3.113	Goddard
8	refinement	PSVS	1.4	Bhattacharya and Montelione
9	chemical shift assignment	AutoAssign	2.30	Zimmerman, Moseley, Kulikowski and Montelione
10	structure solution	PdbStat	5.4	(PdbStat)-Roberto Tejero and Gaetano T. Montelione
11	chemical shift autoassignment	PINE Server	1.0	Bahrami, Markley, Assadi, and Eghbalnia
12	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich

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.