2RN7

NMR solution structure of TnpE protein from Shigella flexneri. Northeast Structural Genomics Target SfR125

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
2	2D 1H-13C HSQC	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
3	3D HNCACB	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
4	3D CBCA(CO)NH	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
5	3D HNCO	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
6	3D HBHA(CO)NH	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
7	3D C(CO)NH	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
8	3D HCCH-TOCSY	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
9	3D HCCH-TOCSY	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
10	2D 1H-15N HSQC	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
11	3D 1H-15N NOESY	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
12	3D 1H-13C NOESY aliph	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
13	3D 1H-13C NOESY arom	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
14	3D HCCH-COSY	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
15	2D 1H-13C HSQC	1mM [U-100% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
16	2D 1H-15N HSQC	1mM [U-7% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293
17	2D 1H-13C HSQC	1mM [U-7% 13C; U-100% 15N] TnpE protein, 10mM TRIS, 500mM sodium chloride, 10mM DTT, 0.01mM Zn+2, 0.01% sodium azide, 1mM benzamidine, 1x protease inhibitor cocktail	90% H2O/10% D2O	.5	7.7	ambient	293

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

NMR Refinement
Method	Details	Software
DGSA-distance geometry simulated annealing, water refinement	Xplor-NIH, CNS	NMRPipe

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	25
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	NMRPipe	nmrpipe_linux	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	collection	VNMR	6.1C	Varian
3	data analysis	AutoStructure	2.1.1	Huang, Tejero, Powers and Montelione
4	structure solution	X-PLOR NIH	2.15.0	Schwieters, Kuszewski, Tjandra and Clore
5	refinement	CNS	1.1	Brunger, Adams, Clore, Gros, Nilges and Read
6	data analysis	Sparky	3.1	Goddard

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.