2ML2

Solution Structure of AlgE6R2 subunit from the Azotobacter vinelandii Mannuronan C5-epimerase

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-1, 25 mM calcium chloride-2, 20 mM HEPES-3, 90 % H2O-4, 10 % D2O-5	90% H2O/10% D2O	0.165	6.9	1 atm	298
2	2D 1H-13C HSQC	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-6, 25 mM calcium chloride-7, 20 mM HEPES-8, 100 % D2O-9	100% D2O	0.165	6.9	1 atm	298
3	3D CBCA(CO)NH	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-1, 25 mM calcium chloride-2, 20 mM HEPES-3, 90 % H2O-4, 10 % D2O-5	90% H2O/10% D2O	0.165	6.9	1 atm	298
4	3D HNCO	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-1, 25 mM calcium chloride-2, 20 mM HEPES-3, 90 % H2O-4, 10 % D2O-5	90% H2O/10% D2O	0.165	6.9	1 atm	298
5	2D 1H-1H TOCSY	1-1.2 mM AlgE6R2-10, 25 mM calcium chloride-11, 20 mM HEPES-12, 100 % D2O-13	100% D2O	0.165	6.9	1 atm	298
6	2D 1H-1H COSY	1-1.2 mM AlgE6R2-10, 25 mM calcium chloride-11, 20 mM HEPES-12, 100 % D2O-13	100% D2O	0.165	6.9	1 atm	298
7	2D 1H-1H TOCSY	1-1.2 mM AlgE6R2-10, 25 mM calcium chloride-11, 20 mM HEPES-12, 100 % D2O-13	100% D2O	0.165	6.9	1 atm	298
8	3D HNCA	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-1, 25 mM calcium chloride-2, 20 mM HEPES-3, 90 % H2O-4, 10 % D2O-5	90% H2O/10% D2O	0.165	6.9	1 atm	298
9	3D HBHA(CO)NH	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-1, 25 mM calcium chloride-2, 20 mM HEPES-3, 90 % H2O-4, 10 % D2O-5	90% H2O/10% D2O	0.165	6.9	1 atm	298
10	3D HN(CO)CA	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-1, 25 mM calcium chloride-2, 20 mM HEPES-3, 90 % H2O-4, 10 % D2O-5	90% H2O/10% D2O	0.165	6.9	1 atm	298
11	3D HCCH-TOCSY	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-6, 25 mM calcium chloride-7, 20 mM HEPES-8, 100 % D2O-9	100% D2O	0.165	6.9	1 atm	298
12	3D HCCH-COSY	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-6, 25 mM calcium chloride-7, 20 mM HEPES-8, 100 % D2O-9	100% D2O	0.165	6.9	1 atm	298
13	3D HBHANH	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-1, 25 mM calcium chloride-2, 20 mM HEPES-3, 90 % H2O-4, 10 % D2O-5	90% H2O/10% D2O	0.165	6.9	1 atm	298
14	3D CBCANH	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-1, 25 mM calcium chloride-2, 20 mM HEPES-3, 90 % H2O-4, 10 % D2O-5	90% H2O/10% D2O	0.165	6.9	1 atm	298
15	3D HN(CA)CO	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-1, 25 mM calcium chloride-2, 20 mM HEPES-3, 90 % H2O-4, 10 % D2O-5	90% H2O/10% D2O	0.165	6.9	1 atm	298
16	3D 1H-13C NOESY aliphatic	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-6, 25 mM calcium chloride-7, 20 mM HEPES-8, 100 % D2O-9	100% D2O	0.165	6.9	1 atm	298
17	3D 1H-15N NOESY	1-1.2 mM [U-98% 13C; U-98% 15N] AlgE6R2-1, 25 mM calcium chloride-2, 20 mM HEPES-3, 90 % H2O-4, 10 % D2O-5	90% H2O/10% D2O	0.165	6.9	1 atm	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	600

NMR Refinement
Method	Details	Software
simulated annealing		XwinNMR

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	XwinNMR	3.5	Bruker Biospin
2	chemical shift assignment	CARA	1.4.1, 1.5.2	Keller and Wuthrich, Krieger E, Koraimann G, Vriend G
3	refinement	CARA	1.4.1, 1.5.2	Keller and Wuthrich, Krieger E, Koraimann G, Vriend G
4	structure solution	CYANA		Guntert, Mumenthaler and Wuthrich
5	refinement	CYANA		Guntert, Mumenthaler and Wuthrich
6	refinement	YASARA		Yasara Biosciences

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.