Experiment: 2IN2

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_15N-separated_NOESY	U-13C/15N; 20mM phosphate buffer; 0.5mM EDTA; 15mM DTT; 0.3% NaN3; 90% H2O, 10% D2O	90% H2O/10% D2O	20 mM	6.5	1 atm	298
2	3D_13C-separated_NOESY	U-13C/15N; 20mM phosphate buffer; 0.5mM EDTA; 15mM DTT; 0.3% NaN3; 90% H2O, 10% D2O	90% H2O/10% D2O	20 mM	6.5	1 atm	298
3	3D_13C-separated_NOESY	U-13C/15N; 20mM phosphate buffer; 0.5mM EDTA; 15mM DTT; NaN3; 99.6% D2O	99.6% D2O	20 mM	6.5	1 atm	298
4	HNHA	U-13C/15N; 20mM phosphate buffer; 0.5mM EDTA; 15mM DTT; 0.3% NaN3; 90% H2O, 10% D2O	90% H2O/10% D2O	20 mM	6.5	1 atm	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	500
2	Varian	INOVA	600
3	Varian	INOVA	800

NMR Refinement
Method	Details	Software
	The structures were calculated with 1515 NOE, 129 phi, 100 psi, 3 chi and 64 hydrogen bond restraints and 29 stereospecific assignments	VNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with acceptable covalent geometry,structures with the least restraint violations,structures with the lowest energy
Conformers Calculated Total Number	300
Conformers Submitted Total Number	20
Representative Model	1 (fewest violations,lowest energy)

Additional NMR Experimental Information
Details	amides involved in hydrogen bonds were identified through H/D exchange

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VNMR	3.1c	Gray, G.
2	processing	NMRPipe	2.3	Delaglio, F.
3	data analysis	NMRView	5.2.2	Johnson, B.
4	structure solution	CYANA	2.1	Guentert, P.
5	refinement	CNS	1.1	Brunger, A.T.

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.