2LV2

Solution NMR structure of C2H2-type Zinc-fingers 4 and 5 from human Insulinoma-associated protein 1 (fragment 424-497), Northeast Structural Genomics Consortium Target HR7614B

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
2	2D 1H-13C HSQC	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
3	3D HNCO	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
4	3D CBCA(CO)NH	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
5	3D HNCACB	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
6	3D 1H-13C NOESY aromatic	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
7	3D 1H-13C NOESY aliphatic	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
8	3D C(CO)NH	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
9	3D HNCA	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
10	3D HN(CO)CA	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
11	3D H(CCO)NH	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
12	3D HCCH-TOCSY	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
13	3D HNHA	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
14	4D CC-NOESY	1.25 mM [U-5% 13C; U-100% 15N] HR7614B.023, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	100% D2O		6.5	ambient	298
15	2D 1H-13C HSQC_CT aliphatic	1.25 mM [U-5% 13C; U-100% 15N] HR7614B.023, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
16	2D 1H-13C HSQC_noCT aliphatic	1.25 mM [U-5% 13C; U-100% 15N] HR7614B.023, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298
17	3D HCCH-TOCSY	1.25 mM [U-5% 13C; U-100% 15N] HR7614B.023, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	100% D2O		6.5	ambient	298
18	3D Nnoesy	0.69 mM [U-100% 13C; U-100% 15N] HR7614B.021, 100 mM NaCl, 5 mM DTT, 0.02 % NaN3, 10 mM Tris-HCl pH 7.5	90% H2O/10% D2O		6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	850
2	Varian	INOVA	600

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	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	data analysis	AutoStructure	2.1	Huang, Tejero, Powers and Montelione
8	refinement	AutoStructure	2.1	Huang, Tejero, Powers and Montelione
9	data analysis	AutoAssign	2.1	Zimmerman, Moseley, Kulikowski and Montelione
10	chemical shift assignment	AutoAssign	2.1	Zimmerman, Moseley, Kulikowski and Montelione
11	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
12	data analysis,peak picking,chemical shift assignment	XEASY		Bartels et al.
13	collection	TopSpin		Bruker Biospin
14	collection	VnmrJ		Varian
15	chemical shift assignment	PINE		Bahrami, Markley, Assadi, and Eghbalnia
16	data analysis	Sparky		Goddard
17	geometry optimization	TALOS+		Shen, Cornilescu, Delaglio and Bax
18	structure validation	PSVS		Bhattacharya, 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.