Experiment: 6ALY

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 500
12	3D 1H-15N TOCSY	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 500
9	3D HBHA(CO)NH	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 500
14	3D H(CCO)NH	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 600
15	3D C(CO)NH	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 600
13	3D 1H-15N NOESY	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 600
11	2D 1H-15N HSQC	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 800
2	3D HNCO	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 800
3	3D HNCA	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 800
4	3D HN(CO)CA	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 800
5	3D CBCA(CO)NH	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 800
6	3D HNCACB	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 800
7	2D 1H-13C HSQC	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 800
10	3D 1H-13C NOESY aromatic	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 600
8	3D 1H-13C NOESY	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	100% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 800
16	3D HCCH-TOCSY	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	100% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 600
17	3D HCCH-COSY	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-13C; U-15N] Med15 ABD2	100% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 600
18	2D 1H-15N HSQC IPAP	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 10 % C12E6/Hexanol, 2.6 mM [U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 800
19	2D 1H-15N HSQC IPAP	20 mM sodium phosphate, 150 mM sodium chloride, 0.1 mM EDTA, 0.1 mM PMSF, 5 mM DTT, 500 uM [U-15N] Med15 ABD2	90% H2O/10% D2O	150 mM	6.5	1 atm	298	Bruker AVANCE III 600

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE III	500
2	Bruker	AVANCE III	600
3	Bruker	AVANCE III	800

NMR Refinement
Method	Details	Software
simulated annealing		X-PLOR NIH

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	chemical shift assignment	NMRView		Johnson, One Moon Scientific
3	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	peak picking	NMRView		Johnson, One Moon Scientific
2	structure calculation	TALOS		Cornilescu, Delaglio and Bax
5	structure calculation	X-PLOR NIH	2.45	Schwieters, Kuszewski, Tjandra and Clore
6	collection	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.

Prepare Data

Validate Data

Deposit Data

Help and Resources

6ALY

Solution structure of yeast Med15 ABD2 residues 277-368