2KJK

Solution structure of the second domain of the listeria protein Lin2157, Northeast Structural Genomics Consortium target Lkr136b

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1.18 mM [U-100% 13C; U-100% 15N] lkr136b, 10 mM DTT, 0.02 % sodium azide, 200 mM sodium chloride, 20 mM sodium acetate, 5 mM calcium chloride	90% H2O/10% D2O	0.2	4.5	ambient	298
2	2D 1H-13C HSQC	1.18 mM [U-100% 13C; U-100% 15N] lkr136b, 10 mM DTT, 0.02 % sodium azide, 200 mM sodium chloride, 20 mM sodium acetate, 5 mM calcium chloride	90% H2O/10% D2O	0.2	4.5	ambient	298
3	3D HNCACB	1.18 mM [U-100% 13C; U-100% 15N] lkr136b, 10 mM DTT, 0.02 % sodium azide, 200 mM sodium chloride, 20 mM sodium acetate, 5 mM calcium chloride	90% H2O/10% D2O	0.2	4.5	ambient	298
4	3D CBCA(CO)NH	1.18 mM [U-100% 13C; U-100% 15N] lkr136b, 10 mM DTT, 0.02 % sodium azide, 200 mM sodium chloride, 20 mM sodium acetate, 5 mM calcium chloride	90% H2O/10% D2O	0.2	4.5	ambient	298
5	3D C(CO)NH	1.18 mM [U-100% 13C; U-100% 15N] lkr136b, 10 mM DTT, 0.02 % sodium azide, 200 mM sodium chloride, 20 mM sodium acetate, 5 mM calcium chloride	90% H2O/10% D2O	0.2	4.5	ambient	298
6	3D 1H-15N NOESY	1.18 mM [U-100% 13C; U-100% 15N] lkr136b, 10 mM DTT, 0.02 % sodium azide, 200 mM sodium chloride, 20 mM sodium acetate, 5 mM calcium chloride	90% H2O/10% D2O	0.2	4.5	ambient	298
7	3D 1H-13C NOESY	1.18 mM [U-100% 13C; U-100% 15N] lkr136b, 10 mM DTT, 0.02 % sodium azide, 200 mM sodium chloride, 20 mM sodium acetate, 5 mM calcium chloride	90% H2O/10% D2O	0.2	4.5	ambient	298
8	2D HSQC/TROSY	1.18 mM [U-100% 13C; U-100% 15N] lkr136b, 10 mM DTT, 0.02 % sodium azide, 200 mM sodium chloride, 20 mM sodium acetate, 5 mM calcium chloride	90% H2O/10% D2O	0.2	4.5	ambient	298
9	2D HSQC/TROSY	1.18 mM [U-100% 13C; U-100% 15N] lkr136b, 4 % pentaethylene glycol monododecyl ether, 200 mM sodium chloride, 0.02 % sodium azide, 10 mM DTT, 20 mM sodium acetate, 5 mM calcium chloride	90% H2O/10% D2O	0.2	4.5	ambient	298

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

NMR Refinement
Method	Details	Software
simulated annealing	structures were first calculated using CYANA's automatic NOE assignment module, which is simulated annealing based. They were then refined in XPLOR-NIH using simulated annealing refinement protocol	CYANA

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	50
Conformers Submitted Total Number	10
Representative Model	1 (fewest violations)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CYANA		Guntert, Mumenthaler and Wuthrich
2	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
3	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	data analysis	NMRView		Johnson, One Moon Scientific

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.