2MKR

Structural Characterization of a Complex Between the Acidic Transactivation Domain of EBNA2 and the Tfb1/p62 subunit of TFIIH.

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.7 mM [U-13C; U-15N] Tfb1, 2.1 mM EBNA2, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
2	2D 1H-15N HSQC	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
3	2D 1H-13C HSQC	0.7 mM [U-13C; U-15N] Tfb1, 2.1 mM EBNA2, 20 mM sodium phosphate, 1 mM DTT	100% D2O	20	6.5	ambient	300
4	2D 1H-13C HSQC	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT-24	100% D2O	20	6.5	ambient	300
5	3D 1H-15N NOESY	0.7 mM [U-15N] Tfb1, 2.1 mM EBNA2, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
6	3D 1H-15N NOESY	0.5 mM [U-100% 15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
7	3D 1H-13C NOESY	0.7 mM [U-13C; U-15N] Tfb1, 2.1 mM EBNA2, 20 mM sodium phosphate, 1 mM DTT	100% D2O	20	6.5	ambient	300
8	3D 1H-13C NOESY	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT-24	100% D2O	20	6.5	ambient	300
9	2D 1H-13C HSQC aromatic	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT-24	100% D2O	20	6.5	ambient	300
10	3D 1H-13C NOESY aromatic	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT-24	100% D2O	20	6.5	ambient	300
11	3D 1H-13C-12C intermolecular NOESY	0.7 mM [U-13C; U-15N] Tfb1, 2.1 mM EBNA2, 20 mM sodium phosphate, 1 mM DTT	100% D2O	20	6.5	ambient	300
12	3D 1H-13C-12C intermolecular NOESY	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT-24	100% D2O	20	6.5	ambient	300
13	3D HNCO	0.7 mM [U-13C; U-15N] Tfb1, 2.1 mM EBNA2, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
14	3D HNCO	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
15	3D HNCACB	0.7 mM [U-13C; U-15N] Tfb1, 2.1 mM EBNA2, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
16	3D HNCACB	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
17	3D HCCH-TOCSY	0.7 mM [U-13C; U-15N] Tfb1, 2.1 mM EBNA2, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
18	3D HCCH-TOCSY	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
19	3D H(CCO)NH	0.7 mM [U-13C; U-15N] Tfb1, 2.1 mM EBNA2, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
20	3D H(CCO)NH	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
21	3D HCCH-COSY	0.7 mM [U-13C; U-15N] Tfb1, 2.1 mM EBNA2, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300
22	3D HCCH-COSY	0.5 mM [U-13C; U-15N] EBNA2, 1.5 mM Tfb1, 20 mM sodium phosphate, 1 mM DTT	90% H2O/10% D2O	20	6.5	ambient	300

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	500
2	Varian	INOVA	600
3	Varian	INOVA	800

NMR Refinement
Method	Details	Software
DGSA-distance geometry 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 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
2	data analysis	CCPNMR		CCPN
3	geometry optimization	TALOS	TALOS-N	Cornilescu, Delaglio and Bax
4	collection	VNMR		Varian
5	refinement	CNS

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.

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2MKR

Structural Characterization of a Complex Between the Acidic Transactivation Domain of EBNA2 and the Tfb1/p62 subunit of TFIIH.