Experiment: 5VF0

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-15N] RAD18, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
2	2D 1H-15N HSQC	0.9 mM [U-100% 13C; U-100% 15N] RAD18, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
3	2D 1H-13C HSQC	0.9 mM [U-100% 13C; U-100% 15N] RAD18, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
4	2D 1H-13C HSQC	0.6 mM [U-100% 13C; U-100% 15N] RAD18, 3 mM Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
5	2D 1H-13C HSQC	3 mM RAD18, 0.6 mM [U-100% 13C; U-100% 15N] Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
6	2D 1H-15N HSQC	0.6 mM [U-100% 13C; U-100% 15N] RAD18, 3 mM Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
7	2D 1H-15N HSQC	3 mM RAD18, 0.6 mM [U-100% 13C; U-100% 15N] Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
8	3D CBCA(CO)NH	0.9 mM [U-100% 13C; U-100% 15N] RAD18, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
9	3D HNCACB	0.9 mM [U-100% 13C; U-100% 15N] RAD18, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
10	3D HBHA(CO)NH	0.9 mM [U-100% 13C; U-100% 15N] RAD18, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
11	3D HCCH-TOCSY	0.9 mM [U-100% 13C; U-100% 15N] RAD18, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
13	3D CBCA(CO)NH	0.6 mM [U-100% 13C; U-100% 15N] RAD18, 3 mM Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
12	3D HNCACB	0.6 mM [U-100% 13C; U-100% 15N] RAD18, 3 mM Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
14	3D HBHA(CO)NH	0.6 mM [U-100% 13C; U-100% 15N] RAD18, 3 mM Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
15	3D HCCH-TOCSY	0.6 mM [U-100% 13C; U-100% 15N] RAD18, 3 mM Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
16	3D CBCA(CO)NH	3 mM RAD18, 0.6 mM [U-100% 13C; U-100% 15N] Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
17	3D HNCACB	3 mM RAD18, 0.6 mM [U-100% 13C; U-100% 15N] Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
18	3D HBHA(CO)NH	3 mM RAD18, 0.6 mM [U-100% 13C; U-100% 15N] Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
19	3D HCCH-TOCSY	3 mM RAD18, 0.6 mM [U-100% 13C; U-100% 15N] Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
20	3D 1H-15N NOESY	0.9 mM [U-15N] RAD18, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
21	3D 1H-15N NOESY	0.6 mM [U-100% 13C; U-100% 15N] RAD18, 3 mM Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
22	3D 1H-15N NOESY	3 mM RAD18, 0.6 mM [U-100% 13C; U-100% 15N] Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
23	3D 1H-13C NOESY	0.9 mM [U-100% 13C; U-100% 15N] RAD18, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
24	3D 1H-13C NOESY	0.6 mM [U-100% 13C; U-100% 15N] RAD18, 3 mM Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
25	3D 1H-13C NOESY	3 mM RAD18, 0.6 mM [U-100% 13C; U-100% 15N] Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
29	3D 15N/13C-FILTERED EDITED NOESY	0.6 mM [U-100% 13C; U-100% 15N] RAD18, 3 mM Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
30	3D 15N/13C-FILTERED EDITED NOESY	3 mM RAD18, 0.6 mM [U-100% 13C; U-100% 15N] Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
31	2D 1H-15N IPAP HSQC	0.2 mM [U-100% 15N] RAD18, 1.0 mM Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2, 5 % Alkyl-polyethylene glycol (C12E5)/n-hexanol mixture	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700
32	2D 1H-15N IPAP HSQC	1.0 mM RAD18, 0.2 mM [U-100% 15N] Ubiquitin, 25 mM MES-Bis-TRIS, 50 mM KCl, 10 uM ZnCl2, 5 % Alkyl-polyethylene glycol (C12E5)/n-hexanol mixture	90% H2O/10% D2O	50 mM KCl mM	6	1 atm	298	Bruker AVANCE III 700

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE III	700

NMR Refinement
Method	Details	Software
simulated annealing		X-PLOR NIH

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin		Bruker Biospin
2	data analysis	Sparky		Goddard
3	peak picking	Sparky		Goddard
6	chemical shift assignment	Sparky		Goddard
5	structure calculation	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
4	refinement	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore

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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5VF0

Solution NMR structure of human RAD18 (198-240) in complex with ubiquitin