2KT7

Solution NMR structure of mucin-binding domain of protein lmo0835 from Listeria monocytogenes, Northeast Structural Genomics Consortium Target LmR64A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	1D 15N T1	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
2	1D 15N T2	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
3	2D 1H-15N HSQC	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
4	2D 1H-13C CT-HSQC aliphatic	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
5	2D 1H-13C CT-HSQC aromatic	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
6	3D HNCO	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
7	3D CBCA(CO)NH	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
8	3D HN(CA)CO	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
9	3D HNCACB	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
10	3D HBHA(CO)NH	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
11	3D (H)CCH-TOCSY aliphatic	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
12	3D (H)CCH-COSY aliphatic	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
13	3D (H)CCH-COSY aromatic	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
14	3D 1H-15N,13C NOESY	0.9 mM [U-13C; U-15N] lmr64a protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
15	2D 1H-15N LR-HSQC (Histidine)	0.9 mM [U-5% 13C; U-15N] lmr64ab protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	6.5	ambient	298
16	2D 1H-13C CT-HSQC methyl	0.9 mM [U-5% 13C; U-15N] lmr64ab protein, 20 mM MES, 100 mM sodium chloride, 5 mM calcium chloride, 0.01 % sodium azide	95% H2O/5% D2O	225	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	The programs CYANA and AutoStructure were used to derive consensus peak assignments followed by iterative refinement with CYANA 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	processing	PROSA	6.4	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	PINE	1.0	Bahrami, Markley, Assadi, and Eghbalnia
7	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
8	structure solution	AutoStructure	2.2.1	Huang, Tejero, Powers and Montelione
9	data analysis	TALOS+	1.2009.0721.18	Shen, Cornilescu, Delaglio and Bax
10	refinement	CNS	1.2.1	Brunger, Adams, Clore, Gros, Nilges and Read
11	validation	PSVS	1.4	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.