2MX9

NMR structure of N-terminal domain from A. ventricosus minor ampullate spidroin (MiSp) at pH 5.5

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.5 mM [U-99% 13C; U-99% 15N] protein, 20 mM sodium chloride, 0.03 % sodium azide, 20 mM [U-99% 2H] sodium acetate, 0.01 tablet/100mL protease inhibitor cocktail	92% H2O/8% D2O	0.04	5.5	ambient	295
2	3D CBCA(CO)NH	0.5 mM [U-99% 13C; U-99% 15N] protein, 20 mM sodium chloride, 0.03 % sodium azide, 20 mM [U-99% 2H] sodium acetate, 0.01 tablet/100mL protease inhibitor cocktail	92% H2O/8% D2O	0.04	5.5	ambient	295
3	3D HNCO	0.5 mM [U-99% 13C; U-99% 15N] protein, 20 mM sodium chloride, 0.03 % sodium azide, 20 mM [U-99% 2H] sodium acetate, 0.01 tablet/100mL protease inhibitor cocktail	92% H2O/8% D2O	0.04	5.5	ambient	295
4	3D HNCA	0.5 mM [U-99% 13C; U-99% 15N] protein, 20 mM sodium chloride, 0.03 % sodium azide, 20 mM [U-99% 2H] sodium acetate, 0.01 tablet/100mL protease inhibitor cocktail	92% H2O/8% D2O	0.04	5.5	ambient	295
5	3D HN(CA)CO	0.5 mM [U-99% 13C; U-99% 15N] protein, 20 mM sodium chloride, 0.03 % sodium azide, 20 mM [U-99% 2H] sodium acetate, 0.01 tablet/100mL protease inhibitor cocktail	92% H2O/8% D2O	0.04	5.5	ambient	295
6	3D 1H-15N NOESY	0.5 mM [U-99% 13C; U-99% 15N] protein, 20 mM sodium chloride, 0.03 % sodium azide, 20 mM [U-99% 2H] sodium acetate, 0.01 tablet/100mL protease inhibitor cocktail	92% H2O/8% D2O	0.04	5.5	ambient	295
7	3D 1H-13C NOESY aromatic	0.5 mM [U-99% 13C; U-99% 15N] protein, 20 mM sodium chloride, 0.03 % sodium azide, 20 mM [U-99% 2H] sodium acetate, 0.01 tablet/100mL protease inhibitor cocktail	92% H2O/8% D2O	0.04	5.5	ambient	295
8	3D 1H-13C NOESY aliphatic	0.5 mM [U-99% 13C; U-99% 15N] protein, 20 mM sodium chloride, 0.03 % sodium azide, 20 mM [U-99% 2H] sodium acetate, 0.01 tablet/100mL protease inhibitor cocktail	92% H2O/8% D2O	0.04	5.5	ambient	295
9	3D 13C,15N filtered 1H-13C NOESY aliphatic	11.3 mg/mL [U-99% 13C; U-99% 15N] protein, 11.3 mg/mL protein, 20 mM sodium chloride, 0.03 % sodium azide, 20 mM [U-2H] sodium acetate, 0.01 tablet/100mL protease inhibitor cocktail	92% H2O/8% D2O	0.04	5.5	ambient	295

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600
2	Bruker	Avance	800

NMR Refinement
Method	Details	Software
simulated annealing, torsion angle dynamics		VNMRJ

NMR Ensemble Information
Conformer Selection Criteria	target function
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	collection	VNMRJ	2.1b	Varian
2	processing	TOPSPIN	3.1	Bruker Biospin
3	collection	TOPSPIN	3.1	Bruker Biospin
4	chemical shift assignment	CARA	1.9.0	Keller and Wuthrich
5	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
6	data analysis	UNIO	2.0.2	T. Herrmann, F. Fiorito, J. Volk
7	peak picking	UNIO	2.0.2	T. Herrmann, F. Fiorito, J. Volk
8	refinement	CNS	1.21	Brunger, Adams, Clore, Gros, Nilges and Read
9	refinement	CYANA		Guntert, Mumenthaler and Wuthrich

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.