Experiment: 2KVQ

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	300 uM [U-100% 13C; U-100% 15N] 15N, 13C -NusG-CTD-1, 300 uM NusE-2, 300 uM NusB-3	90% H2O/10% D2O	50	7.5		310
2	2D 1H-13C HSQC	300 uM [U-100% 13C; U-100% 15N] 15N, 13C -NusG-CTD-1, 300 uM NusE-2, 300 uM NusB-3	90% H2O/10% D2O	50	7.5		310
3	3D 1H-15N NOESY	300 uM [U-100% 13C; U-100% 15N] 15N, 13C -NusG-CTD-1, 300 uM NusE-2, 300 uM NusB-3	90% H2O/10% D2O	50	7.5		310
4	3D 1H-13C NOESY	300 uM [U-100% 13C; U-100% 15N] 15N, 13C -NusG-CTD-1, 300 uM NusE-2, 300 uM NusB-3	90% H2O/10% D2O	50	7.5		310
5	3D 13C filtered(F1)-13C edited NOESY	300 uM [U-100% 13C; U-100% 15N] 15N, 13C -NusG-CTD-1, 300 uM NusE-2, 300 uM NusB-3	90% H2O/10% D2O	50	7.5		310
6	2D 1H-15N HSQC	0.3 mM NusG-CTD-7, 0.3 mM [U-100% 13C; U-100% 15N] 15N, 13C NusE-8, 0.3 mM NusB-9	90% H2O/10% D2O	50	7.5		310
7	2D 1H-13C HSQC	0.3 mM NusG-CTD-7, 0.3 mM [U-100% 13C; U-100% 15N] 15N, 13C NusE-8, 0.3 mM NusB-9	90% H2O/10% D2O	50	7.5		310
8	3D 1H-13C NOESY	0.3 mM NusG-CTD-7, 0.3 mM [U-100% 13C; U-100% 15N] 15N, 13C NusE-8, 0.3 mM NusB-9	90% H2O/10% D2O	50	7.5		310
9	3D 1H-15N NOESY	0.3 mM NusG-CTD-7, 0.3 mM [U-100% 13C; U-100% 15N] 15N, 13C NusE-8, 0.3 mM NusB-9	90% H2O/10% D2O	50	7.5		310
10	3D 13C filtered(F1)-133D 13C filtered(F1)-13C edited NOESY	0.3 mM NusG-CTD-7, 0.3 mM [U-100% 13C; U-100% 15N] 15N, 13C NusE-8, 0.3 mM NusB-9	90% H2O/10% D2O	50	7.5		310
11	3D HNCA	300 uM [U-100% 13C; U-100% 15N; U-80% 2H] 15N, 13C, 2H-NusE-4, 300 uM NusB-5, 300 uM NusG-CTD-6	90% H2O/10% D2O	50	7.5		310
12	3D HNCACB	300 uM [U-100% 13C; U-100% 15N; U-80% 2H] 15N, 13C, 2H-NusE-4, 300 uM NusB-5, 300 uM NusG-CTD-6	90% H2O/10% D2O	50	7.5		310
13	3D HCCH-TOCSY	0.3 mM NusG-CTD-7, 0.3 mM [U-100% 13C; U-100% 15N] 15N, 13C NusE-8, 0.3 mM NusB-9	90% H2O/10% D2O	50	7.5		310
14	3D HCCH-TOCSY	300 uM [U-100% 13C; U-100% 15N] 15N, 13C -NusG-CTD-1, 300 uM NusE-2, 300 uM NusB-3	90% H2O/10% D2O	50	7.5		310

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	700
2	Bruker	AVANCE	800

NMR Refinement
Method	Details	Software
simulated annealing	Structure was calculated using rigid body docking with experimental derived intermolecular distance restraints. The structures of unbound NusG-CTD (pdb 2JVV) and unbound NusE:NusB (pdb 3D3B) were used.	XwinNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	20
Conformers Submitted Total Number	18
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	XwinNMR		Bruker Biospin
2	chemical shift assignment	NMRView		Johnson, One Moon Scientific
3	peak picking	NMRView		Johnson, One Moon Scientific
4	structure solution	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
5	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.