2KW7

Solution NMR Structure of the N-terminal domain of protein PG_0361 from P.gingivalis, Northeast Structural Genomics Consortium Target PgR37A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
2	2D 1H-13C HSQC aliphatic	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
3	3D HNCO	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
4	3D CBCA(CO)NH	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
5	3D HNCACB	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
6	3D HBHA(CO)NH	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
7	3D simutaneous 13C-aromatic,13C-aliphatic,15N edited 1H-1H NOESY	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
8	2D 1H-13C HSQC aromatic	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
9	3D HCCH-COSY aliphatic	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
10	3D HCCH-COSY aromatic	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
11	3D HCCH-TOCSY	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
12	2D 1H-13C HSQC	0.8 mM [U-5% 13C; U-100% 15N] PgR37A-7, 20 mM MES-8, 100 mM sodium chloride-9, 5 mM calcium chloride-10, 0.02 % sodium azide-11, 50 uM DSS-12	95% H2O/5% D2O		6.5	ambient	298
13	2D long-range 1H-15N HSQC for histidine	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298
14	3D HN(CA)CO	0.8 mM [U-100% 13C; U-100% 15N] PgR37A-1, 20 mM MES-2, 100 mM sodium chloride-3, 5 mM calcium chloride-4, 0.02 % sodium azide-5, 50 uM DSS-6	95% H2O/5% D2O		6.5	ambient	298

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

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 and upper limit constraints relaxed by 5%	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	1.2	Brunger, Adams, Clore, Gros, Nilges and Read
2	structure solution	CNS	1.2	Brunger, Adams, Clore, Gros, Nilges and Read
3	geometry optimization	CNS	1.2	Brunger, Adams, Clore, Gros, Nilges and Read
4	structure solution	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
5	structure solution	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
6	structure validation	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
7	data analysis	AutoAssign	2.3	Zimmerman, Moseley, Kulikowski and Montelione
8	chemical shift assignment	AutoAssign	2.3	Zimmerman, Moseley, Kulikowski and Montelione
9	data analysis	CARA	1.8.4	Keller and Wuthrich
10	peak picking	CARA	1.8.4	Keller and Wuthrich
11	chemical shift assignment	CARA	1.8.4	Keller and Wuthrich
12	collection	TopSpin	2.1	Bruker Biospin
13	processing	TopSpin	2.1	Bruker Biospin
14	collection	VnmrJ	2.1B	Varian
15	processing	PROSA	6.4	Guntert
16	data analysis	TALOS+	1.2009.0721.18	Cornilescu, Delaglio and Bax

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.