2M9U

Solution NMR structure of the C-terminal domain (CTD) of Moloney murine leukemia virus integrase, Northeast Structural Genomics Target OR41A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.5 mM [U-100% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
2	2D 1H-13C HSQC	0.5 mM [U-100% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
3	3D HNCO	0.5 mM [U-100% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
4	3D CBCA(CO)NH	0.5 mM [U-100% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
5	3D HNCACB	0.5 mM [U-100% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
6	3D HBHA(CO)NH	0.5 mM [U-100% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
7	3D HCCH-TOCSY	0.5 mM [U-100% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
8	3D 13C-edited_NOESY	0.5 mM [U-100% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
9	3D 1H-15N NOESY	0.5 mM [U-100% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
10	3D 13C-edited_NOESY	0.5 mM [U-100% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
11	2D 1H-15N HSQC	0.5 mM [U-10% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298
12	2D 1H-13C HSQC	0.5 mM [U-10% 13C; U-100% 15N] MLV IN CTD	90% H2O/10% D2O	50mM K-glutamate, 100mM NaCl	6.5	ambient	298

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

NMR Refinement
Method	Details	Software
simulated annealing	The structure was determined using triple resonance NMR spectroscopy. Automated backbone assignments were made using AutoAssign software. Sidechain assignments were completed manually. Initial NOESY assignments for the monomer were made using AutoStructure and CYANA-2.1. The oligomeric state was confirmed to be a monomer by measuring the correlation time using 1D T1 and T2 experiments. A total of 100 structures were calculated and 20 best conformers were then refined in a shell of water using CNS. Final structure quality factors were determined using the PSVS server: ordered residues are defined as (7-18 and 24-61). (a) RMSD (ordered residues) all backbone atoms: 0.6A; all heavy atoms: 0.9A. (b) Ramachandran statistics for all ordered residues: Most favoured region: 87.2%; additionally allowed region: 12.4%; generously allowed region: 0.3% and disallowed region: 0.0%. (c) Procheck scores for all ordered residues (Raw/Z) phi/psi -0.81/-2.87; all dihedral angles: -0.45/-2.66. (d) MolProbity clash score (Raw/Z): 13.47/-0.79. (e) RPF scores for the goodness of fit to NOESY data:- Recall: 0.888; precision: 0.968; F-measure: 0.926; DP score: 0.821	AutoAssign

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	chemical shift assignment	AutoAssign	2.2.1	Zimmerman, Moseley, Kulikowski and Montelione
2	chemical shift assignment	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
3	structure solution	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
4	geometry optimization	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
5	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
6	geometry optimization	CNS	2.0.6	Brunger, Adams, Clore, Gros, Nilges and Read
7	refinement	CNS	2.0.6	Brunger, Adams, Clore, Gros, Nilges and Read
8	peak picking	Sparky	2.1	Goddard

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.