2HTG | pdb_00002htg

Structural and functional characterization of TM VII of the NHE1 isoform of the Na+/H+ exchanger

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D NOESY	~ 1 mM peptide synthetic source 1 (unlabeled), ~ 75 mM dodecylphosphocholine, 1.0 mM DSS	90% H2O/10% D2O	~ 75 mM Na+/DPC-	4.8	ambient	303
2	2D TOCSY	~ 1 mM peptide synthetic source 1 (unlabeled), ~ 75 mM dodecylphosphocholine, 1.0 mM DSS	90% H2O/10% D2O	~ 75 mM Na+/DPC-	4.8	ambient	303
3	13C-natural abundance HSQC	~ 1 mM peptide synthetic source 1 (unlabeled), ~ 75 mM dodecylphosphocholine, 1.0 mM DSS	90% H2O/10% D2O	~ 75 mM Na+/DPC-	4.8	ambient	303
4	2D NOESY	~ 1 mM peptide synthetic source 2 (6 specific 15N labels), ~ 75 mM dodecylphosphocholine, 1.0 mM DSS	90% H2O/10% D2O	~ 75 mM Na+/DPC-	4.8	ambient	303
5	2D TOCSY	~ 1 mM peptide synthetic source 2 (6 specific 15N labels), ~ 75 mM dodecylphosphocholine, 1.0 mM DSS	90% H2O/10% D2O	~ 75 mM Na+/DPC-	4.8	ambient	303
6	13C-natural abundance HSQC	~ 1 mM peptide synthetic source 2 (6 specific 15N labels), ~ 75 mM dodecylphosphocholine, 1.0 mM DSS	90% H2O/10% D2O	~ 75 mM Na+/DPC-	4.8	ambient	303

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	800
2	Varian	INOVA	500

NMR Refinement
Method	Details	Software
simulated annealing; torsion angle dynamics	1311 unique NOE restraints were used; 25 dihedral angle restraints. All inter-residue restraints were made ambiguous and calculations were carried out using 2 identical polypeptide chains	BioPack

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	120
Conformers Submitted Total Number	66
Representative Model	1 (n/a)

Additional NMR Experimental Information
Details	15N labels were used to assist in assignments. 2D homonuclear NOESY data from labeled and unlabeled samples were pooled for structure calculation.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	BioPack		Varian
2	processing	NMRPipe	2005.319.14.56	Delaglio et al.
3	data analysis	Sparky	3.110	Goddard & Kneller
4	structure solution	XPLOR-NIH	2.13	Schwieters et al.
5	refinement	XPLOR-NIH	2.13	Schwieters et al.

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 R01GM157729.