2KY4

Solution NMR structure of the PBS linker domain of phycobilisome linker polypeptide from Anabaena sp. Northeast Structural Genomics Consortium Target NsR123E

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
2	2D 1H-13C CT-HSQC ali	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
3	2D 1H-13C CT-HSQC aro	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
4	2D 1H-13C CT-HSQC (methyl)	0.975 mM [U-5% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
5	3D HNCO	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
6	3D HN(CA)CO	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
7	3D CBCA(CO)NH	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
8	3D HNCACB	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
9	3D HCCH-TOCSY	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
10	3D HCCH-COSY ali	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
11	3D HCCH-COSY aro	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
12	3D HBHA(CO)NH	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
13	13C/15N-NOESY	0.95 mM [U-100% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298
14	2D J-modulation 1H-15N HSQC	0.664 mM [U-5% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS, 4 % by volume poly ethylene glycol	82% H2O/18% D2O	117.5	6.5	ambient	298
15	2D J-modulation 1H-15N HSQC	0.851 mM [U-5% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS, 7 % by volume poly acrylamide gel	83% H2O/17% D2O	117.5	6.5	ambient	298
16	2D J-modulation 1H-15N HSQC	0.975 mM [U-5% 13C; U-100% 15N] protein, 100 mM sodium chloride, 5 mM calcium chloride, 10 mM DTT, 20 mM MES, 0.02 % by volume NaN3, 50 uM DSS	90% H2O/10% D2O	117.5	6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	750
2	Varian	INOVA	600
3	Bruker	AVANCE	900

NMR Refinement
Method	Details	Software
simulated annealing	Structure determination was performed iteratively with CYANA v3.0 using NOE-based constraints, PHI and PSI dihedral angle constraints from TALOS+, and RDC constraints. 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	14
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VnmrJ	2.1B	Varian
2	processing	NMRPipe	2007.030.16.06	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	processing	PROSA	6.4	Guntert
4	data analysis	XEASY		Bartels et al.
5	chemical shift assignment	AutoAssign		Zimmerman, Moseley, Kulikowski and Montelione
6	structure solution	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
7	structure solution	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
8	data analysis	TALOS+	1.2009.0721.18	Shen, Cornilescu, Delaglio and Bax
9	chemical shift assignment	CARA	1.8.4	Keller and Wuthrich
10	peak picking	CARA	1.8.4	Keller and Wuthrich
11	data analysis	CARA	1.8.4	Keller and Wuthrich
12	refinement	CNS	1.2.1	Brunger, Adams, Clore, Gros, Nilges and Read
13	validation	PSVS	1.3	Bhattacharya and Montelione
14	data analysis	NMRDraw	3.0	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
15	refinement	MOLMOL		Koradi, Billeter and Wuthrich
16	data analysis	TopSpin	2.1	Bruker Biospin

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.