Experiment: 2MW4

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	2.5 mM [U-100% 15N] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
2	2D 1H-13C HSQC	2.5 mM [U-100% 13C; U-100% 15N] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
3	3D HNCACB	2.5 mM [U-100% 13C; U-100% 15N] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
4	3D HNCO	2.5 mM [U-100% 13C; U-100% 15N] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
5	3D HCCCONH	2.5 mM [U-100% 13C; U-100% 15N] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
6	3D CCCONH	2.5 mM [U-100% 13C; U-100% 15N] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
7	3D HCCH-TOCSY	2.5 mM [U-100% 13C; U-100% 15N] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
8	3D 1H-13C NOESY aliphatic	2.5 mM [U-100% 13C; U-100% 15N] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
9	3D 1H-13C NOESY aromatic	2 mM [U-100% 15N] protein, 2 mM [U-100% 13C] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
10	3D 1H-13C NOESY 15N-edited inter	2 mM [U-100% 15N] protein, 2 mM [U-100% 13C] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
11	3D 1H-13C HMQC-NOESY-15N-1H-TROSY	2 mM [U-100% 15N] protein, 2 mM [U-100% 13C] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
12	3D 1H-13C NOESY J-resolved	2 mM [U-100% 15N] protein, 2 mM [U-100% 13C] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298
13	3D 1H-15N NOESY	2.5 mM [U-100% 15N] protein, 50 mM L-arginine, 50 mM L-glutamate, 1 mM DTT	95% H2O/5% D2O	0	7.0	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	900
2	Bruker	AVANCE	800
3	Bruker	AVANCE	700
4	Bruker	AVANCE	600
5	Bruker	AVANCE	500

NMR Refinement
Method	Details	Software
simulated annealing, energy minimization	SIMULATED ANNEALING WITH TORSION ANGLE DYNAMICS, REFINEMENT IN EXPLICIT WATER	CYANA

NMR Ensemble Information
Conformer Selection Criteria	target function
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CYANA	3.9	Guntert, Mumenthaler and Wuthrich
2	structure solution	CNS	1.1	Brunger, Adams, Clore, Gros, Nilges and Read
3	structure solution	ARIA	1.2	Linge, O'Donoghue and Nilges
4	refinement	ARIA	1.2	Linge, O'Donoghue and Nilges
5	collection	TopSpin	2.1	Bruker Biospin
6	processing	TopSpin	2.1	Bruker Biospin
7	chemical shift assignment	Sparky	3.114	Goddard
8	data analysis	Sparky	3.114	Goddard
9	peak picking	Sparky	3.114	Goddard

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.