2EZW

Solution structure of the docking and dimerization domain of the type I alpha regulatory subunit of protein kinase A (RIalpha D/D)

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D NOESY	R1a(12-61) at 1.2-1.6 mM dimer, 50mM sodium acetate, 150mM sodium chloride, pH 4.0, 90% H2O, 10% D2O	90% H2O/10% D2O	50mM sodium acetate, 150mM sodium chloride	4.	1 atm	310
2	2D 1H-15N HSQC, amide proton exchange	15N-enriched R1a(12-61)	5% H2O, 95% D2O	50mM sodium acetate, 150mM sodium chloride	4.	1 atm	310
3	3D 1H-15N HSQC NOESY	15N-enriched R1a(12-61), 90% H2O, 10% D2O	90% H2O/10% D2O	50mM sodium acetate, 150mM sodium chloride	4.	1 atm	310
4	3D HNHA	15N-enriched R1a(12-61), 90% H2O, 10% D2O	90% H2O/10% D2O	50mM sodium acetate, 150mM sodium chloride	4.	1 atm	310
5	3D 13C-edited HMQC-NOESY	13C/15N-enriched R1a(12-61)	5% H2O, 95% D2O	50mM sodium acetate, 150mM sodium chloride	4.	1 atm	310
6	3D 13C-edited(w2) 12C-filtered(w1) 13C-filtered(w3) NOESY	asymmetrically enriched 13C/15N-12C/14N R1a(12-61)	5% H2O,95% D2O	50mM sodium acetate, 150mM sodium chloride	4.	1 atm	310

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	DMX	500
2	Bruker	DRX	600

NMR Refinement
Method	Details	Software
simulated annealing	The structures are based on a total of 435 NOE-derived distance, 139 backbone dihedral and 13 hydrogen bond restraints per monomer	FELIX

NMR Ensemble Information
Conformer Selection Criteria
Conformers Calculated Total Number
Conformers Submitted Total Number	18
Representative Model	1 (n/a)

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

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	FELIX	95.0	Molecular Simulations Inc.
2	structure solution	X-PLOR	3.851	Brunger, A.T.
3	refinement	X-PLOR	3.851	Brunger, A.T.

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.