2KJ6

NMR Solution Structure of a Tubulin folding cofactor B obtained from Arabidopsis thaliana: Northeast Structural Genomics Consortium target AR3436A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D 1H-15N NOESY	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
2	3D 1H-13Caliphatic NOESY	0.78 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
3	3D 1H-13C aromatic NOESY	0.78 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
4	2D 1H-15N HSQC	0.78 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
5	2D 1H-13C HSQC	0.78 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
6	2D 1H-15N HSQC	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
7	2D 1H-13C HSQC	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
8	3D HBHA(CO)NH	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
9	3D CBCA(CO)NH	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
10	(4,3)D GFT-HNNCABCA	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
11	(4,3)D GFT-CABCA(CO)NHN	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
12	3D HNCO	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
13	3D HNCA	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
14	3D HNCACB	0.95 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
15	3D HNHA	0.56 mM [U-10% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
16	3D H(CCO)NH	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
17	3D HCCH-TOCSY	0.4 mM [U-100% 13C; U-100% 15N] AR3436A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	800
2	Varian	INOVA	600
3	Varian	INOVA	500

NMR Refinement
Method	Details	Software
simulated annealing	The structure was obtained using triple resonance NMR spectroscopy. GFT-NMR experiments were used for backbone assignments. Automated NOESY assignments were made using AUTOSTRUCTURE and CYANA-2.1. Dihedral angle constraints were obtained from TALOS. The assignments were validated using AVS software. Final structure factors determined using PSVS: ordered residues: 29-33, 15-21, 89-94, 57-62, 39-50, 79-82. Ramachandran statistics for ordered residues: Most favored region: 84.4%, additionally favored region: 15.6%, generously aloowed: 0.1%. Procheck scores for ordered residues (RAW/Z): Phi/psi -0.51/-1.69, all -0.33/-1.95, MolProbity clash scores (RAW/Z) - 15.41/-1.12. RPF scores for the goodness fit to NOESY data: Recall-0.993, Precision-0.878, F-measure-0.932, final dp score-0.756.	CNS

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	140
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	CNS	2.0.6	Brunger, Adams, Clore, Gros, Nilges and Read
2	structure solution	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
3	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
4	chemical shift assignment	AutoAssign	2.4.0	Zimmerman, Moseley, Kulikowski and Montelione

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.