2K1S

Solution NMR structure of the folded C-terminal fragment of YiaD from Escherichia coli. Northeast Structural Genomics Consortium target ER553.

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
2	2D 1H-13C HSQC	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
3	2D 1H-13C HSQC	1.3 mM [U-5% 13C; U-100% 15N] YiaD 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
4	3D 1H-15N NOESY	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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-13C NOESY	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
6	3D 1H-13C NOESY	1.9 mM [U-100% 13C; U-100% 15N] YiaD protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide	100% D2O	0.1	6.5	ambient	293
7	3D HNCO	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
8	3D HNCA	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
9	3D HN(CO)CA	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
10	3D HNCACB	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
11	3D CBCA(CO)NH	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
12	3D C(CO)NH	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
13	3D HBHA(CO)NH	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
14	3D H(CCO)NH	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
15	3D HCCH-TOCSY	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
16	3D HCCH-COSY	1.9 mM [U-100% 13C; U-100% 15N] YiaD 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
17	4D CC NOESY	1.9 mM [U-100% 13C; U-100% 15N] YiaD protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide	100% D2O	0.1	6.5	ambient	293

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

NMR Refinement
Method	Details	Software
simulated annealing	Xplor-NIH and CNS water refinement	NMRPipe

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
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	linux9	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	collection	VNMR	6.1C	Varian
3	collection	TopSpin	2.1	Bruker Biospin
4	data analysis	AutoStructure	2.1.1	Huang, Tejero, Powers and Montelione
5	structure solution	X-PLOR NIH	2.15.0	Schwieters, Kuszewski, Tjandra and Clore
6	refinement	CNS	1.1	Brunger, Adams, Clore, Gros, Nilges and Read
7	data analysis	Sparky	3.113	Goddard
8	structure solution	PSVS	1.3	Bhattacharya and Montelione
9	chemical shift assignment	AutoAssign		Zimmerman, Moseley, Kulikowski 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.