2M9A

Solution NMR Structure of E3 ubiquitin-protein ligase ZFP91 from Homo sapiens, Northeast Structural Genomics Consortium (NESG) Target HR7784A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
2	2D 1H-13C HSQC aromatic	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
3	3D CBCA(CO)NH	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
4	3D HNCACB	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
5	3D HNCO	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
6	3D H(CCO)NH	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
7	3D HCCH-TOCSY	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
8	3D 1H-15N NOESY	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
9	3D 1H-13C NOESY aliphatic	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
10	3D 1H-13C NOESY aromatic	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
11	2D CB(CGCD)HD	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
12	2D CB(CGCDCE)HE	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
13	2D 1H-15N J-modulation HSQC	0.66 mM [U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 4.2 % C12E5, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298
14	2D 1H-15N J-modulation HSQC	0.66 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS, 5 % positively charged polyacrylamide gel	95% H2O/5% D2O	100	6.5	ambient	298
15	2D 1H-15N J-modulation HSQC	1.17 mM [U-100% 13C; U-100% 15N] HR7784A, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 0.02 % sodium azide, 50 uM DSS	95% H2O/5% D2O	100	6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600

NMR Refinement
Method	Details	Software
simulated annealing		CYANA

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	50
Conformers Submitted Total Number	20
Representative Model	1 (fewest violations)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
2	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	chemical shift assignment	Sparky	3.113	Goddard
4	peak picking	Sparky	3.113	Goddard
5	refinement	CNS	1.3	Brunger, Adams, Clore, Gros, Nilges and Read
6	data analysis	PSVS	1.4	Bhattacharya and Montelione

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.