2M83

Solution structure of the carbohydrate binding module of the muscle glycogen-targeting subunit of Protein Phosphatase-1

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.8 mM [U-99% 15N] GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	90% H2O/10% D2O	70	6.5	ambient	298
2	3D HNCACB	0.8 mM [U-99% 13C; U-99% 15N] GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	90% H2O/10% D2O	70	6.5	ambient	298
3	3D CBCA(CO)NH	0.8 mM [U-99% 13C; U-99% 15N] GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	90% H2O/10% D2O	70	6.5	ambient	298
4	3D HNCA	0.8 mM [U-99% 13C; U-99% 15N] GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	90% H2O/10% D2O	70	6.5	ambient	298
5	3D (H)CC(CO)NH	0.8 mM [U-99% 13C; U-99% 15N] GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	90% H2O/10% D2O	70	6.5	ambient	298
6	3D HBHA(CO)NH	0.8 mM [U-99% 13C; U-99% 15N] GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	90% H2O/10% D2O	70	6.5	ambient	298
7	3D HCCH-TOCSY	0.8 mM [U-99% 13C; U-99% 15N] GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	90% H2O/10% D2O	70	6.5	ambient	298
8	2D 1H-1H NOESY	0.8 mM GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	100% D2O	70	6.5	ambient	298
9	2D 1H-1H TOCSY	0.8 mM GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	100% D2O	70	6.5	ambient	298
10	2D 1H-1H COSY	0.8 mM GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	100% D2O	70	6.5	ambient	298
11	3D 1H-15N NOESY	0.8 mM [U-99% 15N] GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	90% H2O/10% D2O	70	6.5	ambient	298
12	3D 1H-13C NOESY	0.8 mM [U-99% 13C; U-99% 15N] GM CBM21, 20 mM sodium phosphate, 50 mM sodium chloride, 10 mM DTT	90% H2O/10% D2O	70	6.5	ambient	298

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

NMR Refinement
Method	Details	Software
simulated annealing	RECOORD scripts were used (Nederveen et al. Proteins. 2005.)	CNS

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	refinement	CNS		Brunger A. T. et.al.
2	chemical shift assignment	CARA		Keller and Wuthrich
3	peak picking	CARA		Keller and Wuthrich
4	collection	TopSpin		Bruker Biospin
5	processing	TopSpin		Bruker Biospin
6	structure solution	CYANA_2.1		Guntert, Mumenthaler and Wuthrich

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.