Experiment: 2ROC

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D 1H-15N NOESY	0.5mM [U-100% 13C; U-100% 15N] Mcl-1; 0.5mM Puma; 95% H2O/5% D2O	95% H2O/5% D2O	120 mM	6.7	ambient	298
2	3D 1H-13C NOESY	0.5mM [U-100% 13C; U-100% 15N] Mcl-1; 0.5mM Puma; 95% H2O/5% D2O	95% H2O/5% D2O	120 mM	6.7	ambient	298
3	3D HNHA	0.5mM [U-100% 13C; U-100% 15N] Mcl-1; 0.5mM Puma; 95% H2O/5% D2O	95% H2O/5% D2O	120 mM	6.7	ambient	298
4	3D 1H-15N NOESY	0.5mM Mcl-1; 0.5mM [U-100% 13C; U-100% 15N] Puma; 95% H2O/5% D2O	95% H2O/5% D2O	120 mM	6.7	ambient	298
5	3D 1H-13C NOESY	0.5mM Mcl-1; 0.5mM [U-100% 13C; U-100% 15N] Puma; 95% H2O/5% D2O	95% H2O/5% D2O	120 mM	6.7	ambient	298
6	3D HNHA	0.5mM Mcl-1; 0.5mM [U-100% 13C; U-100% 15N] Puma; 95% H2O/5% D2O	95% H2O/5% D2O	120 mM	6.7	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	500
2	Bruker	DRX	600
3	Bruker	AVANCE	800

NMR Refinement
Method	Details	Software
DGSA-distance geometry simulated annealing	Structures were calculated on the basis of 3667 total constraints. 2943 of these constraints were NOE-derived distances of which 197 were intermolecular NOEs; 348 dihedral angle restraints and 188 hydrogen bonds	TopSpin

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	256
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average)

Additional NMR Experimental Information
Details	standard heteronuclear methods were used to assign resonances and determine the constraints

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	TopSpin	1.3	Bruker Biospin
2	data analysis	XEASY	1.3	Bartels et al.
3	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
4	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.