Experiment: 2K31

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 15N] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
2	2D 1H-13C HSQC	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
3	2D 1H-1H NOESY	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
4	3D HNCO	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N; 80% 2H] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
5	3D HNCA	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N; 80% 2H] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
6	3D HNCACB	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N; 80% 2H] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
7	3D CBCA(CO)NH	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N; 80% 2H] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
8	3D HCCH-COSY	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
9	3D HN(COCA)CB	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N; 80% 2H] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
10	3D HCCH-TOCSY	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
11	3D 1H-15N NOESY	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
12	3D 1H-13C NOESY	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	100% D2O	150	7	ambient atm	310
13	3D HN(CO)CA	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 13C; U-100% 15N; 80% 2H] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	90% H2O/10% D2O	150	7	ambient atm	310
14	2D 1H-15N HSQC	0.5 - 1.2 mM GUANOSINE-3',5'-MONOPHOSPHATE, 0.5 - 1.2 mM [U-100% 15N] mPDE5_GAFA, 150 mM sodium chloride, 1 mM PMSF, 1-5 mM DTT, 0.1 mM EDTA, 25 mM sodium phosphate	100% D2O	150	7	ambient atm	310

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DMX	500
2	Bruker	DMX	600
3	Varian	INOVA	600
4	Varian	INOVA	800
5	Varian	INOVA	900

NMR Refinement
Method	Details	Software
simulated annealing		CNS

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CNS	1.1	Brunger, Adams, Clore, Gros, Nilges and Read
2	structure solution	CYANA	2.0	Guntert, Mumenthaler and Wuthrich
3	processing	NMRDraw		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	chemical shift assignment	NMRView		Johnson, One Moon Scientific
5	peak picking	NMRView		Johnson, One Moon Scientific
6	data analysis	NMRView		Johnson, One Moon Scientific
7	data analysis	ModelFree		Palmer
8	data analysis	TALOS		Cornilescu, Delaglio and Bax
9	refinement	CYANA	2.0	Guntert, Mumenthaler and Wuthrich

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.