2KSF

Backbone structure of the membrane domain of E. coli histidine kinase receptor KdpD, Center for Structures of Membrane Proteins (CSMP) target 4312C

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.5 mM DSS, 20 mM Mes-BisTris, 100 mM 1-myristoyl-2-hydroxy-sn-glycero-3-[phospho-rac-(1-glycerol)], 0.3 mM [U-99% 13C; U-99% 15N] Protein	95% H2O/5% D2O		6.0	ambient	318
2	3D HNCO	0.5 mM DSS, 20 mM Mes-BisTris, 100 mM 1-myristoyl-2-hydroxy-sn-glycero-3-[phospho-rac-(1-glycerol)], 0.3 mM [U-99% 13C; U-99% 15N] Protein	95% H2O/5% D2O		6.0	ambient	318
3	3D HNCA	0.5 mM DSS, 20 mM Mes-BisTris, 100 mM 1-myristoyl-2-hydroxy-sn-glycero-3-[phospho-rac-(1-glycerol)], 0.3 mM [U-99% 13C; U-99% 15N] Protein	95% H2O/5% D2O		6.0	ambient	318
4	3D HNCACB	0.5 mM DSS, 20 mM Mes-BisTris, 100 mM 1-myristoyl-2-hydroxy-sn-glycero-3-[phospho-rac-(1-glycerol)], 0.3 mM [U-99% 13C; U-99% 15N] Protein	95% H2O/5% D2O		6.0	ambient	318
5	3D HN(CO)CA	0.5 mM DSS, 20 mM Mes-BisTris, 100 mM 1-myristoyl-2-hydroxy-sn-glycero-3-[phospho-rac-(1-glycerol)], 0.3 mM [U-99% 13C; U-99% 15N] Protein	95% H2O/5% D2O		6.0	ambient	318
6	3D 1H-15N NOESY	0.5 mM DSS, 20 mM Mes-BisTris, 100 mM 1-myristoyl-2-hydroxy-sn-glycero-3-[phospho-rac-(1-glycerol)], 0.3 mM [U-99% 13C; U-99% 15N] Protein	95% H2O/5% D2O		6.0	ambient	318

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	700

NMR Refinement
Method	Details	Software
torsion angle dynamics, simulated annealing		CYANA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CYANA	1.0.6	Guntert, Mumenthaler and Wuthrich
2	data analysis	CARA	2K.2	Keller and Wuthrich
3	peak picking	CARA	2K.2	Keller and Wuthrich
4	refinement	CNS	1.1	Brunger, Adams, Clore, Gros, Nilges and Read

RCSB PDB Core Operations are funded by the U.S. National Science Foundation (DBI-2321666), 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.