Experiment: 2K2W

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
2	2D 1H-13C HSQC	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
3	3D CBCA(CO)NH	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
4	3D HNCACB	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
5	3D HBHA(CO)NH	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
6	3D HNCO	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
7	3D HCACO	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
8	3D H(CCO)NH	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
9	3D C(CO)NH	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
10	3D HNCACB	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
11	3D HCCH-COSY	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
12	3D CCH-COSY	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
13	3D CCH-TOCSY	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
14	3D 1H-15N NOESY	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
15	3D 1H-15N TOCSY	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
16	3D 1H-13C NOESY aliphatic	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
17	2D HBCBCGCDHD	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288
18	3D 1H-13C AROMATIC	20 mM sodium phosphate, 50 mM sodium chloride, 50 mM glutamic acid, 50 mM arginine, 0.3-0.5 mM [U-100% 13C; U-100% 15N] protein	93% H2O/7% D2O	0.1	6.7	ambient	288

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	700
2	Bruker	AVANCE	600

NMR Refinement
Method	Details	Software
simulated annealing		CYANA

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	structure solution	CYANA		Guntert, Mumenthaler and Wuthrich
2	refinement	Amber	8.0	Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... and Kollm
3	data analysis	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	data analysis	NMRView		Johnson, One Moon Scientific
5	data analysis	TALOS		Wishart

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.