6QH2

Solution NMR ensemble for a chimeric KH-S1 domain construct of exosomal polynucleotide phosphrylase at 298K compiled using the CoMAND method

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D HNCACB	0.4 mM [U-99% 13C; U-99% 15N] KH-S1, 50 mM potassium phosphate, 100 mM sodium chloride	90% H2O/10% D2O	125 mM	7.5	ambient atm	298	Bruker AVANCE III 600
2	3D CBCA(CO)NH	0.4 mM [U-99% 13C; U-99% 15N] KH-S1, 50 mM potassium phosphate, 100 mM sodium chloride	90% H2O/10% D2O	125 mM	7.5	ambient atm	298	Bruker AVANCE III 600
3	3D C(CO)NH	0.4 mM [U-99% 13C; U-99% 15N] KH-S1, 50 mM potassium phosphate, 100 mM sodium chloride	90% H2O/10% D2O	125 mM	7.5	ambient atm	298	Bruker AVANCE III 600
4	3D HCCH-TOCSY	0.4 mM [U-99% 13C; U-99% 15N] KH-S1, 50 mM potassium phosphate, 100 mM sodium chloride	90% H2O/10% D2O	125 mM	7.5	ambient atm	298	Bruker AVANCE III 600
5	3D CNH-NOESY	0.4 mM [U-99% 13C; U-99% 15N] KH-S1, 50 mM potassium phosphate, 100 mM sodium chloride	90% H2O/10% D2O	125 mM	7.5	ambient atm	298	Bruker AVANCE III 800

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

NMR Refinement
Method	Details	Software
R-factor based frame picking	frames picked from unrestrained MD simulations to minimise overall CNH-NOESY based R-Factors	CoMAND

NMR Ensemble Information
Conformer Selection Criteria	back calculated data agree with experimental NOESY spectrum
Conformers Calculated Total Number	10000
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin		Bruker Biospin
3	chemical shift assignment	Sparky		Goddard
4	structure calculation	Shine		Riss & Coles
2	structure calculation	Rosetta	3.6	Das & Baker
5	refinement	NAMD	2.12	University of Illinois
6	refinement	CoMAND		in house

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.