5W72

Impact of IR active probes on PDZ3 and its ligand binding studied by NMR and X-ray crystallography

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker AVANCE III HD 800
2	2D 1H-1H NOESY	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker AVANCE III HD 700
3	3D 1H-15N NOESY	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker AVANCE III HD 800
4	3D 1H-13C NOESY	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker DRX 800
5	2D 13C-15N df-NOESY	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker AVANCE III HD 700
7	3D 13C edited 13C-15N df-NOESY	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker AVANCE III HD 700
6	3D 15N edited 13C-15N df-NOESY	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker AVANCE III HD 700
8	3D HNCACB	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker AVANCE III HD 600
11	3D HNCO	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker AVANCE III HD 800
10	3D HNHA	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker AVANCE III HD 800
9	3D HCCCONH	0.5 mM [U-98% 13C; U-98% 15N] PDZ3, 50 mM sodium phosphate	90% H2O/10% D2O	0 mM	6.8	1 atm	298	Bruker AVANCE III HD 600

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE III HD	700
2	Bruker	DRX	800
3	Bruker	AVANCE III HD	800
4	Bruker	AVANCE III HD	600

NMR Refinement
Method	Details	Software
torsion angle dynamics	CYANA	Sparky

NMR Ensemble Information
Conformer Selection Criteria	target function
Conformers Calculated Total Number	400
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure calculation	CYANA	3.97	Guentert, P.
2	chemical shift assignment	Sparky		Goddard
3	collection	TopSpin		Bruker Biospin
4	processing	TopSpin		Bruker Biospin

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.