2KK8

NMR Solution Structure of a Putative Uncharacterized Protein obtained from Arabidopsis thaliana: Northeast Structural Genomics Consortium target AR3449A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D 1H-15N NOESY	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
2	3D 1H-13C aliphatic NOESY	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
3	3D 1H-13C aromatic NOESY	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
4	2D 1H-15N HSQC	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
5	2D 1H-13C HSQC	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
6	2D 1H-15N HSQC	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
7	2D 1H-13C HSQC	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
8	3D HBHA(CO)NH	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
9	3D CBCA(CO)NH	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
10	(4,3)D GFT-HNNCABCA	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
11	(4,3)D GFT-CABCA(CO)NHN	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
12	3D HNCO	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
13	3D HNCA	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
14	3D HNCACB	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
15	3D H(CCO)NH	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
16	3D HCCH-TOCSY	1.11 mM [U-100% 13C; U-100% 15N] AR3449A	90% H2O/10% D2O	5mM CaCl2, 10mM NaCl	6.5	ambient	293
17	3D HNHA	1.22 mM [U-10% 13C; U-100% 15N] AR3449A	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 obteined from TALOS. The assignments were validated using AVS software. FInal structure factors determined using PSVS: ordered residues: alpha helices- 33.44, 67-70, beta strands- 22-27, 11-17, 78-82, 51-55, 58-59. Ramachandran statistics for ordered residues: 89.1% most favoured regions, 10.9% additionally favoured regions. Procheck scores for ordered residues (RAW/Z): phi/psi -0.43/-1.38, all -0.18/-1.06, MolProbity clashscores- 21.46/-2.16. RPF score for goodness fit to the NOESY data: Recall-0.998, Precision-0.93, final dp-score-0.869	CNS

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	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.