2P6J | pdb_00002p6j

Full-sequence computational design and solution structure of a thermostable protein variant

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	aliphatic 3D_13C-separated_NOESY	Uniform (random) labeling with 13C, 15N: U-13C, U-15N, 95% H2O, 5% D2O	95% H2O/5% D2O	50 mM sodium phosphate	5.5	1 atm	293
2	aromatic 3D_13C-separated_NOESY	Uniform (random) labeling with 13C, 15N: U-13C, U-15N, 95% H2O, 5% D2O	95% H2O/5% D2O	50 mM sodium phosphate	5.5	1 atm	293
3	3D_15N-separated_NOESY	Uniform (random) labeling with 13C, 15N: U-13C, U-15N, 95% H2O, 5% D2O	95% H2O/5% D2O	50 mM sodium phosphate	5.5	1 atm	293

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

NMR Refinement
Method	Details	Software
simulated annealing, distance geometry	The structures are based on a total of 1321 restraints: 1245 are NOE-derived distance constraints, 57 are dihedral angle restraints and 19 distance restraints are from hydrogen bonds.	VNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with acceptable covalent geometry,structures with the least restraint violations
Conformers Calculated Total Number	100
Conformers Submitted Total Number	43
Representative Model	1 (closest to the average)

Additional NMR Experimental Information
Details	The structure was determined using triple-resonance NMR spectroscopy. Hydrogen bond restraints were derived from protection factors measured by H/D exchange. Torsion angle restraints were derived from J-coupling data (HNHA experiment) and from TALOS predictions.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VNMR	6.1C	varian
2	processing	NMRPipe		Delaglio, F. et al.
3	data analysis	NMRView	4.0	Johnson, B.A. et al.
4	refinement	ARIA	1.2	Nilges, M. et al.
5	structure solution	ARIA	1.2	Nilges, M. 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.