Experiment: 2LXC

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1 mM Get5, 2 mM [U-100% 13C; U-100% 15N] Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
2	3D 1H-15N NOESY	1 mM Get5, 2 mM [U-100% 13C; U-100% 15N] Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
3	3D 1H-15N TOCSY	1 mM Get5, 2 mM [U-100% 13C; U-100% 15N] Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
4	2D 1H-13C HSQC aliphatic	1 mM Get5, 2 mM [U-100% 13C; U-100% 15N] Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
5	2D 1H-13C HSQC aromatic	1 mM Get5, 2 mM [U-100% 13C; U-100% 15N] Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
6	2D 1H-15N HSQC	1 mM [U-100% 13C; U-100% 15N] Get5, 2 mM Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
7	3D HNCACB	1 mM [U-100% 13C; U-100% 15N] Get5, 2 mM Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
8	3D HNCO	1 mM [U-100% 13C; U-100% 15N] Get5, 2 mM Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
9	3D HCCH-TOCSY	1 mM [U-100% 13C; U-100% 15N] Get5, 2 mM Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
10	3D HCCH-TOCSY	1 mM [U-100% 13C; U-100% 15N] Get5, 2 mM Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
11	3D 1H-15N NOESY	1 mM [U-100% 13C; U-100% 15N] Get5, 2 mM Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
12	3D 1H-13C NOESY aliphatic	1 mM [U-100% 13C; U-100% 15N] Get5, 2 mM Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide	90% H2O/10% D2O	0	6.1	ambient	310
13	2D 1H-15N HSQC	1 mM Get5, 2 mM [U-100% 13C; U-100% 15N] Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide, 3.5 % polyacrylamide	90% H2O/10% D2O	0	6.1	ambient	310
14	2D 1H-15N HSQC	1 mM [U-100% 13C; U-100% 15N] Get5, 2 mM Sgt2, 10 mM Bis-Tris, 50 mM L-arginine, 50 mM L-glutamic acid, 0.02 % sodium azide, 3.5 % polyacrylamide	90% H2O/10% D2O	0	6.1	ambient	310

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600
2	Bruker	AVANCE	800

NMR Refinement
Method	Details	Software
simulated annealing, experimentally driven docking	STRUCTURES OF ASYMMETRICALLY LABELED COMPLEX (SAMPLE_1 AND SAMPLE_3) WERE USED TO DETERMINE THE SEPERATE STRUCTURES OF GET5 OR SGT2, IN THEIR COMPLEX STATES. STRUCTURES ARE REFINED IN EXPLICIT WATER. AMBIGUOUS INTERACTION RESTRAINTS, RESIDUAL DIPOLAR COUPLINGS AND INTERMOLECULAR NOES WERE USED TO DOCK THE STRUCTURES OF GET5 AND SGT2, DETERMINED IN THEIR COMPLEX STATES. STRUCTURES ARE SELECTED FROM THE BEST SCORING CLUSTER BY HADDOCK SCORES AND REFINED IN WATER.	VnmrJ

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VnmrJ		Varian
2	chemical shift assignment	Analysis		CCPN
3	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	data analysis	TALOS+		Cornilescu, Delaglio and Bax
5	structure solution	ARIA	2.3	Linge, O'Donoghue and Nilges
6	refinement	ARIA	2.3	Linge, O'Donoghue and Nilges
7	structure solution	HADDOCK	2.1	Alexandre Bonvin
8	collection	TopSpin		Bruker Biospin
9	data analysis	PALES		Zweckstetter and Bax

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.