Experiment: 2NBI

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D HNCA	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
2	3D CBCA(CO)NH	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
3	2D 1H-15N HSQC	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
4	1H-15N NOESY HSQC	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
5	1H-15N TOCSY HSQC	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
6	3D HNCO	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
7	HACACO	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
8	3D HCCH-TOCSY	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
9	2D 1H-1H TOCSY	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
10	2D 1H-1H NOESY	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
11	3D HBHA(CO)NH	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
12	2D 1H-13C HSQC	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298
13	2D 1H-15N HSQC	0.8 mM [U-97% 13C, U-98% 15N] Pleuralin, formerly HEP200	95% H2O/5% D2O		6.5		298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	600
2	Bruker	DRX	800

NMR Refinement
Method	Details	Software
simulated annealing, na	Structures were refined by using data imputation (. Cano, C., Brunner, Baskaran, K., K. Elsner, R., Munte, C. E. and Kalbitzer, H. R. (2009) Protein structure calculation with data imputation: the use of substitute restraints. J. Biomol. NMR 45, 397-411) and refinement in explicut water (Linge, J.P., Williams, M.A., Spronk, C.A.E.M., Bonvin, A.M.J.J. and Nilges, M. (2003). Refinement of protein structures in explicit solvent. Proteins 50, 496-506)., The structure of the PSCD4-domain has been simulated using the experimental restraints obtained from the NMR spectra. The other four domains have been modeled transfering the restraints of the PSCD4-domain for those residues that were kept identical in the sequences. Additional substitute restraints of each domain (Noe distances, hydrogen bonds and dihedral angles) have been obtained using the PERMOL routine	AUREMOL

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	2000
Conformers Submitted Total Number	1
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	The structure has been determined combining experimental restraints (J-coupling, Noe-distances, Hydrogen Bonds and dihedral angles) with additional substitute restraints obtained by homology modelling between the fourth and the other domains

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	peak picking	AUREMOL	2.4.1 beta	Bruker Biospin
2	data analysis	AUREMOL	2.4.1 beta	Bruker Biospin
3	processing	AUREMOL	2.4.1 beta	Bruker Biospin
4	structure solution	CNS	1.21	Brunger, Adams, Clore, Gros, Nilges and Read
5	collection	Topspin	3.1	Bruker Biospin
6	processing	Topspin	3.1	Bruker Biospin
7	data analysis	Topspin	3.1	Bruker Biospin
8	refinement	CNS
9	refinement	AUREMOL

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.