2JU2

Minimized mean solution structure of the acyl carrier protein domain from module 2 of 6-deoxyerythronolide B synthase (DEBS)

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D 1H-15N NOESY	0.8-1 mM [U-13C; U-15N] acyl carrier protein, 0.05 % sodium azide, 30 mM sodium phosphate	90% H2O/10% D2O	30	5.5	ambient	288
2	3D 1H-13C/15N NOESY	0.8-1 mM [U-13C; U-15N] acyl carrier protein, 0.05 % sodium azide, 30 mM sodium phosphate	90% H2O/10% D2O	30	5.5	ambient	288
3	3D HCCH-TOCSY	0.8-1 mM [U-13C; U-15N] acyl carrier protein, 0.05 % sodium azide, 30 mM sodium phosphate	90% H2O/10% D2O	30	5.5	ambient	288
4	3D HNHA	0.8-1 mM [U-13C; U-15N] acyl carrier protein, 0.05 % sodium azide, 30 mM sodium phosphate	90% H2O/10% D2O	30	5.5	ambient	288
5	2D 1H-15N HSQC	0.8-1 mM [U-13C; U-15N] acyl carrier protein, 0.05 % sodium azide, 30 mM sodium phosphate	90% H2O/10% D2O	30	5.5	ambient	288
6	3D HNCACB	0.8-1 mM [U-13C; U-15N] acyl carrier protein, 0.05 % sodium azide, 30 mM sodium phosphate	90% H2O/10% D2O	30	5.5	ambient	288
7	3D C(CO)NH	0.8-1 mM [U-13C; U-15N] acyl carrier protein, 0.05 % sodium azide, 30 mM sodium phosphate	90% H2O/10% D2O	30	5.5	ambient	288

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	800
2	Varian	INOVA	500
3	Varian	INOVA	600

NMR Refinement
Method	Details	Software
simulated annealing, torsion angle dynamics	The standard simulated annealing protocol included as part of DYANA software was used to calculate a total of 50 structures, of which 30 structures with the lowest target function values were chosen for the solution structure ensemble. The helical regions of these 30 structures were superimposed and the mean structure was obtained using MOLMOL software, then subjected to the standard variable target function minimization implemented in DYANA. The resulting minimized mean structure is deposited here. The ensemble of 30 structures is deposited in the PDB entry 2ju1.	DYANA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	50
Conformers Submitted Total Number	1
Representative Model	1 (minimized average structure)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	DYANA	1.5	Guntert, P. et al.
2	refinement	DYANA	1.5	Guntert, P. et al.

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.