2KPT

Solution NMR structure of the N-terminal domain of cg2496 protein from Corynebacterium glutamicum. Northeast Structural Genomics Consortium Target CgR26A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
2	2D 1H-13C HSQC aliphatic	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
3	2D 1H-13C CT-HSQC aliphatic	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
4	2D 1H-13C CT-HSQC aromatic	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
5	3D HNCO	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
6	3D CBCA(CO)NH	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
7	3D HNCACB	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
8	3D HN(CA)CO	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
9	3D HBHA(CO)NH	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
10	3D (H)CCH-COSY aliphatic	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
11	3D (H)CCH-COSY aromatic	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
12	3D (H)CCH-TOCSY aliphatic	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
13	1D 15N T1	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
14	1D 15N T2	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
15	3D 1H-15N/13C NOESY	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
16	2D (HB)CB(CGCDCE)HE aromatic	0.9 mM [U-100% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
17	2D 1H-13C CT-HSQC methyl 28ms	0.9 mM [U-5% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298
18	2D 1H-13C CT-HSQC methyl 56ms	0.9 mM [U-5% 13C; U-100% 15N] cgr26a, 20 mM MES, 200 mM sodium chloride, 5 mM calcium chloride, 50 uM DSS, 0.02 % sodium azide	90% H2O/10% D2O	225	6.5	ambient	298

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

NMR Refinement
Method	Details	Software
simulated annealing	Structure determination was performed iteratively with CYANA v2.1 using NOE-based constraints and PHI and PSI dihedral angle constraints from TALOS+. The 20 conformers out of 100 with the lowest target function were further refined by simulated annealing in explicit water bath using the program CNS with PARAM19 force field.	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	collection	TopSpin	2.1	Bruker Biospin
3	processing	TopSpin	2.1	Bruker Biospin
4	processing	PROSA	6.4	Guntert
5	data analysis	XEASY	1.3.13	Bartels et al.
6	data analysis	CARA	1.8.4	Keller and Wuthrich
7	chemical shift assignment	CARA	1.8.4	Keller and Wuthrich
8	peak picking	CARA	1.8.4	Keller and Wuthrich
9	chemical shift assignment	PINE	1.0	Bahrami, Markley, Assadi, and Eghbalnia
10	data analysis	TALOS+	1.2009.0721.18	Shen, Cornilescu, Delaglio and Bax
11	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
12	structure solution	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
13	structure validation	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
14	structure validation	PSVS	1.4	Bhattacharya and Montelione
15	refinement	CNS	1.2.1	Brunger, Adams, Clore, Gros, Nilges and Read

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.