2LSO

Solution NMR Structure of the Globular Domain of Human Histone H1x, Northeast Structural Genomics Consortium (NESG) Target HR7057A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
2	2D 1H-13C CT-HSQC aliphatic	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
3	3D HNCO	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
4	3D CBCA(CO)NH	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
5	3D HNCACB	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
6	3D (H)CCH-TOCSY aliphatic	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
7	3D simutaneous 13C-aromatic,13C-aliphatic,15N edited 1H-1H NOESY	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
8	2D 1H-13C CT-HSQC aromatic	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
9	3D (H)C(CCO)NH-TOCSY	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
10	3D H(CCCO)NH-TOCSY	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
11	3D HBHA(CO)NH	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
12	3D HN(CA)CO	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
13	3D HCCH-COSY aliphatic	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
14	3D HCCH-COSY aromatic	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
15	2D 1H-13C CT-HSQC methyl	0.6 mM [5% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
16	2D J-resolved 1H-15N HSQC	0.6 mM [5% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
17	2D J-resolved 1H-15N HSQC	0.3 mM [5% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide, 4 % C12E5 PEG, 4 % hexanol	90% H2O/10% D2O		6.5	ambient	298
18	1D 15N T1	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298
19	1D 15N T2	0.9 mM [U-100% 13C; U-100% 15N] HR7057A.011, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O		6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	750
2	Varian	INOVA	600
3	Varian	INOVA	600

NMR Refinement
Method	Details	Software
simulated annealing	Structure determination was performed by running CYANA and ASDP in parallel using NOE-based constraints and PHI and PSI dihedral angle constraints from TALOS+. Consensus peak assignments were selected and used in iterative refinement with CYANA, with RDC constraints added at later stages. The 20 conformers out of 100 with the lowest target function were further refined by simulated annealing in explicit water bath using the program CNS with PARAM19 force field	CNS

NMR Ensemble Information
Conformer Selection Criteria	target function
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	1.2	Brunger, Adams, Clore, Gros, Nilges and Read
2	structure solution	CNS	1.2	Brunger, Adams, Clore, Gros, Nilges and Read
3	geometry optimization	CNS	1.2	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	ASDP	1.0	Huang, Tejero, Powers and Montelione
8	refinement	ASDP	1.0	Huang, Tejero, Powers and Montelione
9	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
10	data analysis	XEASY	1.3.13	Bartels et al.
11	collection	VnmrJ	2.2D	Varian
12	chemical shift assignment	PINE	1.0	Bahrami, Markley, Assadi, and Eghbalnia
13	data analysis	Sparky		Goddard
14	geometry optimization	TALOS+		Shen, Cornilescu, Delaglio and Bax
15	structure validation	PSVS	1.4	Bhattacharya, Montelione
16	data analysis	CARA	1.8.4	Keller and Wuthrich
17	chemical shift assignment	CARA	1.8.4	Keller and Wuthrich
18	peak picking	CARA	1.8.4	Keller and Wuthrich

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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2LSO

Solution NMR Structure of the Globular Domain of Human Histone H1x, Northeast Structural Genomics Consortium (NESG) Target HR7057A