2M8X

Restrained CS-Rosetta Solution NMR structure of the CARDB domain of PF1109 from Pyrococcus furiosus. Northeast Structural Genomics Consortium target PfR193A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
2	2D 1H-13C HSQC	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
3	3D 1H-15N NOESY	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
4	3D 1H-13C NOESY aliphatic	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
5	3D 1H-13C NOESY aromatic	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
6	2D 1H-13C HSQC high resolution (L/V methyl stereoassignment)	0.43 mM [U-5% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
7	3D HNCO	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
8	3D HN(CA)CO	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
9	3D HN(CO)CA	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
10	3D HNCA	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
11	3D CBCA(CO)NH	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
12	3D HNCACB	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
13	3D HBHA(CO)NH	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
14	3D HCCH-TOCSY	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
15	3D HCCH-COSY	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
16	3D CCH-TOCSY	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
17	3D HNHA	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
18	2D 1H-15N hetNOE	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298
19	1D 15N T1 and T2	0.65 mM [U-100% 13C; U-100% 15N] PfR193A, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 10 % D2O, 90 % H2O	90% H2O/10% D2O	0.2	6.5	1 atm	298

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

NMR Refinement
Method	Details	Software
	Structures were calculated by Restrained CS-Rosetta method using NOE-based distance and dihedral angle restraints from the original CNS-water refined structure calculations	TopSpin

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	10000
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	THE FIRST REPRESENTATIVE STRUCTURE IN THE ENSEMBLE IS THE MEDOID STRUCTURE.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin	2.1	Bruker Biospin
2	data analysis	TopSpin	2.1	Bruker Biospin
3	processing	NMRPipe	2.3	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	data analysis	Sparky	3.112	Goddard
5	peak picking	Sparky	3.112	Goddard
6	chemical shift assignment	PINE	1.0	Bahrami, Markley, Assadi, and Eghbalnia
7	structure solution	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
8	rpf analysis	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
9	structure quality analysis	PSVS	1.3	Bhattacharya and Montelione
10	structure quality analysis	MolProbity	3.15	Richardson
11	pdb coordinate analysis	PdbStat	5.9	Tejero and Montelione
12	dihedral angle constraints	TALOS	plus	Cornilescu, Delaglio and Bax
13	refinement	Rosetta	3	Rohl, Strauss, Misura, and Baker

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.

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2M8X

Restrained CS-Rosetta Solution NMR structure of the CARDB domain of PF1109 from Pyrococcus furiosus. Northeast Structural Genomics Consortium target PfR193A