2KAN

Solution NMR structure of ubiquitin-like domain of Arabidopsis thaliana protein At2g32350. Northeast Structural Genomics Consortium target AR3433A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
2	2D 1H-13C HSQC (ali)	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
3	2D 1H-13C CT-HSQC (ali)	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
4	2D 1H-13C HSQC (aro)	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
5	3D HNCO	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
6	3D CBCA(CO)NH	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
7	3D HNCACB	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
8	3D HBHA(CO)NH	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
9	3D HCCH-COSY (ali)	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
10	3D HCCH-TOCSY	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
11	(4,3)D GFT HCCH-COSY (aro)	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
12	3D HCCH-COSY (aro)	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
13	3D 1H-15N,13Cali,13Caro NOESY	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
14	2D 1H-15N LR-HSQC (His)	1.09 mM [U-100% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298
15	2D 1H-13C CT-HSQC (ali)	1.15 mM [U-5% 13C; U-100% 15N] ar3433a protein, 50 uM DSS, 200 mM sodium chloride, 20 mM MES, 5 mM calcium cloride, 10 mM DTT, 0.02 % sodium azide, 1 X protease inhibitors	90% H2O/10% D2O	215	6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	500
2	Varian	INOVA	750

NMR Refinement
Method	Details	Software
simulated annealing		VnmrJ

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

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VnmrJ	2.1B	Varian
2	processing	PROSA	6.0.2	Guntert
3	chemical shift assignment	CARA	1.8.4	Keller and Wuthrich
4	data analysis	CARA	1.8.4	Keller and Wuthrich
5	peak picking	CARA	1.8.4	Keller and Wuthrich
6	chemical shift assignment	AutoAssign	1.15.1	Zimmerman, Moseley, Kulikowski and Montelione
7	data analysis	TALOS	2003.027.13.05	Cornilescu, Delaglio and Bax
8	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
9	structure solution	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
10	structure validation	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
11	refinement	CNS	1.2	Brunger, Adams, Clore, Gros, Nilges and Read
12	structure validation	PSVS	1.3	Bhattacharya 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.