1Q10

Ensemble of 40 Structures of the Dimeric Mutant of the B1 Domain of Streptococcal Protein G

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_13C-separated_NOESY	1.7mM (in monomer) U-15N,13C, 50mM phosphate buffer, 0.02% NaN3	92.5% H2O, 7.5% D2O	50mM sodium phosphate buffer	5.5	ambient	298
2	3D_15N-separated_NOESY	1.7mM (in monomer) U-15N,13C, 50mM phosphate buffer, 0.02% NaN3	92.5% H2O, 7.5% D2O	50mM sodium phosphate buffer	5.5	ambient	298
3	3D_13C-separated/12C-filtered_NOESY	0.85mM (in monomer) U-15N,13C, 0.85mM (in monomer) unlabeled, 50mM phosphate buffer (pH 5.5), 0.02 % NaN3	92.5% H2O, 7.5% D2O	50mM sodium phosphate buffer	5.5	ambient	298
4	3D_13C-separated/12C-filtered_NOESY	0.85mM (in monomer) U-15N,13C, 0.85mM (in monomer) unlabeled, 50mM phosphate buffer, 0.02% NaN3	100% D2O	50mM sodium phosphate buffer	5.5	ambient	298
5	2D NOESY	1.7mM (in monomer) unlabeled, 50mM phosphate buffer, 0.02% NaN3	92.5% H2O, 7.5% D2O	50mM sodium phosphate buffer	5.5	ambient	298
6	2D NOESY	1.7mM (in monomer) unlabeled, 50mM phosphate buffer, 0.02% NaN3	100% D2O	50mM sodium phosphate buffer	5.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DRX	800
2	Bruker	DMX	750
3	Bruker	DRX	600
4	Bruker	DMX	600
5	Bruker	DMX	500

NMR Refinement
Method	Details	Software
simulated annealing	The structures are based on a total of 1311 constraints per monomeric subunit: 1035 NOE-derived distance constraints, 72 hydrogen bond distance constraints, 148 dihedral angle constraints, and 56 residual N-H dipolar coupling constraints.	XwinNMR

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy,target function
Conformers Calculated Total Number	100
Conformers Submitted Total Number	40
Representative Model	1 (minimized average structure)

Additional NMR Experimental Information
Details	The structure was determined using triple-resonance NMR spectroscopy.

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	XwinNMR	2.6 and 3.1	Bruker
2	processing	NMRPipe	2.2	Delaglio
3	data analysis	PIPP	4.3.2	Garrett
4	structure solution	CNS	1.1	Brunger
5	refinement	CNS	1.1	Brunger

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.