1XSB

Structure of the nudix enzyme AP4A hydrolase from homo sapiens (E63A mutant) in complex with ATP. No ATP restraints included

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	C HSQC-NOESY	1.0mM human AP4A hydrolase	20mM MgCl2, 20mM imidazole, pH 6.5, 90% H2O 10% D2O, 10mM DTT, 1mM EDTA, 1.5mM ATP	52.5mM	6.5	ambient	293
2	3D_15N-separated_NOESY	1.0mM human AP4A hydrolase	20mM MgCl2, 20mM imidazole, pH 6.5, 90% H2O 10% D2O, 10mM DTT, 1mM EDTA, 1.5mM ATP	52.5mM	6.5	ambient	293
3	13C HSQC-NOESY	1.0mM human AP4A hydrolase	20mM MgCl2, 20mM imidazole, pH 6.5, 100% D2O, 10mM DTT, 1mM EDTA, 1.5mM ATP	52.5mM	6.5	ambient	293
4	13C NOESY-HSQC aromatic region	1.0mM human AP4A hydrolase	20mM MgCl2, 20mM imidazole, pH 6.5, 100% D2O, 10mM DTT, 1mM EDTA, 1.5mM ATP	52.5mM	6.5	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600
2	Varian	INOVA	500

NMR Refinement
Method	Details	Software
CANDID xplor-NIH RAMA potential. Refine against CA & CB shifts.		CYANA

NMR Ensemble Information
Conformer Selection Criteria	target function
Conformers Calculated Total Number	100
Conformers Submitted Total Number	39
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	Residues 1 and 2 are cganged from GP to Alanine in the calculations. These are non native from precission cleavage site and are unstructured. ATP constraints are not included. Different calculation protocol is used for this and deposited separately (lower temp, slower cooling).

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CYANA	1.0.7	P Guntert
2	refinement	XPLOR-NIH	2.9.1

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.