2LMD

Minimal Constraints Solution NMR Structure of Prospero Homeobox protein 1 from Homo sapiens, Northeast Structural Genomics Consortium Target HR4660B

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.96 mM [U-100% 13C; U-100% 15N] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298
2	2D 1H-13C HSQC	0.96 mM [U-100% 13C; U-100% 15N] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298
3	3D HNCO	0.2 mM [U-13C; U-15N; U-2H] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298
4	3D CBCA(CO)NH	0.2 mM [U-13C; U-15N; U-2H] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298
5	3D HNCACB	0.2 mM [U-13C; U-15N; U-2H] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298
6	3D 1H-13C arom NOESY	0.96 mM [U-100% 13C; U-100% 15N] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298
7	3D 1H-15N NOESY	0.96 mM [U-100% 13C; U-100% 15N] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298
8	3D 1H-13C-13C HSQC_NOESY_HSQC	0.2 mM [U-13C; U-15N; U-2H] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298
9	3D 1H-13C NOESY	0.96 mM [U-100% 13C; U-100% 15N] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298
10	3D 1H-13C NOESY	0.2 mM [U-13C; U-15N; U-2H] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298
11	2D 1H-13C HSQC	0.86 mM [5% 13C; U-100% 15N] HR4660B, 200 mM sodium chloride, 10 mM DTT, 5 mM Calcium Chloride, 0.02 % sodium azide, 20 mM MES	95% H2O/5% D2O		6.5	ambient	298

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

NMR Refinement
Method	Details	Software
molecular dynamics	MD steps: 200 HEAT-1000 HOT 100 COOL, PARAM 19 timestep 0.004 ns heat, 0.004 ns hot, 0.001 ns cool, 0.3 weight rdc	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	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
2	structure solution	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
3	geometry optimization	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
4	refinement	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
5	geometry optimization	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
6	structure solution	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
7	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
8	collection	TopSpin		Bruker Biospin
9	chemical shift assignment	PINE		Bahrami, Markley, Assadi, and Eghbalnia
10	data analysis	Sparky		Goddard
11	geometry optimization	TALOS+		Shen, Cornilescu, Delaglio and Bax
12	geometry optimization	PALES		PALES (Zweckstetter, Bax)
13	data analysis	PSVS		Bhattacharya and Montelione

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.