2KCX

Solution NMR Structure of Kazal-1 Domain of Human Follistatin-related protein 3 (FSTL-3). Northeast Structural Genomics Target HR6186A.

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
2	2D 1H-13C HSQC	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
3	3D CBCA(CO)NH	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
4	3D 1H-15N NOESY	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
5	3D 1H-13C NOESY	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
6	3D HNCACB	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
7	3D HNCO	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
8	3D HBHA(CO)NH	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
9	3D HCCH-TOCSY	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
10	3D HCCH-COSY	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
11	3D CCH-TOCSY	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
12	3D HNCA	0.6 mM [U-100% 13C; U-100% 15N] FSTL-3-1, 20 mM MES-2, 200 mM sodium chloride-3	90% H2O/10% D2O	0.2	6.5	ambient	298
13	2D 1H-15N HET_NOE	0.2 mM [U-100% 15N] FSTL-3-4, 20 mM MES-5, 200 mM sodium chloride-6	90% H2O/10% D2O	0.2	6.5	ambient	298
14	2D N15 T1	0.2 mM [U-100% 15N] FSTL-3-4, 20 mM MES-5, 200 mM sodium chloride-6	90% H2O/10% D2O	0.2	6.5	ambient	298
15	2D N15 T2 cpmg	0.2 mM [U-100% 15N] FSTL-3-4, 20 mM MES-5, 200 mM sodium chloride-6	90% H2O/10% D2O	0.2	6.5	ambient	298
16	2D 1H-15N HSQC	0.2 mM [U-100% 15N] FSTL-3-4, 20 mM MES-5, 200 mM sodium chloride-6	90% H2O/10% D2O	0.2	6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	800

NMR Refinement
Method	Details	Software
torsion angle dynamics	MONOMER BY GEL FILTRATION CHROMATOGRAPHY/LIGHT SCATTERING AND BY NMR. T1/T2(CPMG) (MS) = 516.1/136.2, TAUC = 3.94(NS). CONSISTENT WITH MOLECULAR WEIGHT. STRUCTURE DETERMINED BY TRIPLE RESONANCE NMR SPECTROSCOPY. NOESY ASSIGNMENTS BY CYANA2.1. 20 OF 100 STRUCTURES LOWEST TARGET FUNCTION SELECTED WITH CYANA-2.1. SELECTED MODELS ARE FURTHER REFINED USING CNS IN EXPLICIT WATER SHELL (NILGES PROTOCOL WITH PARAM19). ASSIGNMENT STATS (EXCLUDING C-TERM TAG): BACKBONE 98.10%, SIDECHAIN 88.50%, AROMATIC (SC) 75.00%. STRUCTURE BASED ON 912 NOE, 65 DIHE. MAX NOE VIOLATION: 0.19 A (1MODEL); MAX DIHE VIOLATION: 5.4 DEG. 6 TOTAL CLOSE CONTACTS PER 20 MODELS. STRUCTURE QUALITY FACTOR (PSVS 1.3): ORDERED RESIDUES RANGES: 4-9, 13-27, 33-34, 37-58, 61-69, 72-73 FOR [S(PHI)+S(PSI)] > 1.8. SECONDARY STRUCTURE - ALPHA HELICES: 50-60, BETA STRANDS: 13-18, 21-26, 46-47, 39-41, 66-69. RMSD 1.3 BACKBONE, 1.8 ALL HEAVY ATOMS. RAMA. DISTRIBUTION: 92.1/7.8/0.1/0.0. PROCHECK (PSI-PHI): -0.34/-1.02 (RAW/Z), PROCHECK (ALL): -0.33/-1.95 (RAW/Z), MOLPROBITY CLASH: 19.69/-1.85 (RAW/Z). RPF SCORES ALL ASSIGNED RESIDUES (FIT OF NOESY PEAKLISTS TO STRUCTURE): RECALL: 0.99, PRECISION: 0.85, F-MEASURE: 0.91, DP-SCORE: 0.82.	CYANA

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	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
2	geometry optimization	TALOS		Cornilescu, Delaglio and Bax
3	structure solution	PSVS	1.3	Bhattacharya and Montelione
4	refinement	CNS	1.2	Brunger, Adams, Clore, Gros, Nilges and Read
5	processing	NMRPipe	2008	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
6	data analysis	Sparky	2.113	Goddard
7	collection	TopSpin	2.1	Bruker Biospin
8	chemical shift assignment	PINE		Bahrami, Markley, Assadi, and Eghbalnia
9	visualization	MOLMOL		Koradi, Billeter and Wuthrich
10	refinement	Procheck		Laskowski, MacArthur, Smith, Jones, Hutchinson, Morris, Moss and Tho
11	refinement	MolProbity		Richardson
12	data analysis	PdbStat		Tejero, 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.