2LEQ

Chemical Shift Assignment and Solution Structure of ChR145 from Cytophaga Hutchinsonii, Northeast Structural Genomics Consortium Target ChR145

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
2	2D 1H-13C HSQC aromatic	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
3	3D CBCA(CO)NH	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
4	3D C(CO)NH	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
5	3D HNCO	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
6	3D HNCACB	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
7	3D HBHA(CO)NH	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
8	3D H(CCO)NH	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
9	3D HCCH-TOCSY	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
10	3D 1H-15N NOESY	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
11	3D 1H-13C NOESY aliphatic	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
12	3D 1H-13C NOESY aromatic	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES	95% H2O/5% D2O	200	6.5	ambient	298
13	NH J-Modulation	0.57 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES, 12.5 mg Pf1 phage	95% H2O/5% D2O	200	6.5	ambient	298
14	NH J-Modulation	0.57 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES, 4 % C12E5/Hexanol	95% H2O/5% D2O	200	6.5	ambient	298
15	NH J-Modulation	0.85 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride-25, 20 mM MES-26, 5 % Polyacrylamide-27	95% H2O/5% D2O	200	6.5	ambient	298
16	NC J-Modulation	0.57 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES, 12.5 mg Pf1 phage	95% H2O/5% D2O	200	6.5	ambient	298
17	NC J-Modulation	0.57 mM [U-100% 13C; U-100% 15N] ChR145, 0.2 % sodium azide, 5 mM DTT, 5 mM CaCl2, 200 mM sodium chloride, 20 mM MES, 4 % C12E5/Hexanol	95% H2O/5% D2O	200	6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600

NMR Refinement
Method	Details	Software
simulated annealing		CYANA

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	50
Conformers Submitted Total Number	10
Representative Model	1 (fewest violations)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
2	data analysis	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	peak picking	Sparky		Goddard
4	chemical shift assignment	Sparky		Goddard
5	refinement	X-PLOR NIH	2.18	Schwieters, Kuszewski, Tjandra and Clore
6	validation	PSVS		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.