2OI3

NMR Structure Analysis of the Hematopoetic Cell Kinase SH3 Domain complexed with an artificial high affinity ligand (PD1)

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D_13C-separated_NOESY	1.3mM Hck-SH3 U-13C, U-15N: 1.3mM PD1, 20mM KPO4, 20mM NaCl, pH 6.7, 93% H2O, 7% D2O	93% H2O/7% D2O	20mM KPO4, 20mM NaCl	6.7	1 atm	298
2	3D_15N-separated_NOESY	1.3mM Hck-SH3 U-13C, U-15N: 1.3mM PD1, 20mM KPO4, 20mM NaCl, pH 6.7, 93% H2O, 7% D2O	93% H2O/7% D2O	20mM KPO4, 20mM NaCl	6.7	1 atm	298
3	3D_13C/15N-filtered/edited NOESY	1.3mM Hck-SH3 U-13C, U-15N: 1.3mM PD1, 20mM KPO4, 20mM NaCl, pH 6.7, 93% H2O, 7% D2O	93% H2O/7% D2O	20mM KPO4, 20mM NaCl	6.7	1 atm	298
4	2D_13C/15N-double filtered NOESY	1.3mM Hck-SH3 U-13C, U-15N: 1.3mM PD1, 20mM KPO4, 20mM NaCl, pH 6.7, 93% H2O, 7% D2O	93% H2O/7% D2O	20mM KPO4, 20mM NaCl	6.7	1 atm	298

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

NMR Refinement
Method	Details	Software
Simulated annealing with generalized born solvent model and NMR distance constraints		NMRPipe

NMR Ensemble Information
Conformer Selection Criteria	Minimized average structure
Conformers Calculated Total Number	20
Conformers Submitted Total Number	1
Representative Model	1 (minimized average structure)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	NMRPipe	2.3	Delaglio
2	data analysis	CARA/NEASY	1.3.1	Keller
3	refinement	RADAR	0.9b	Herrmann
4	refinement	Amber	8.0	Case
5	processing	VnmrJ	1.1d	Varian, Inc.
6	structure solution	CYANA	1.1	Guentert

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.