7A2D

Structure-function analyses of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localisation to the cell division site

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
14	3D HCCH-TOCSY	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
2	3D HN(CO)CA	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
3	3D HNCA	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
4	3D CBCA(CO)NH	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
5	3D HNCACB	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
6	3D HNCO	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
7	3D HN(CA)CO	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
8	3D HCCH-TOCSY	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
10	3D 1H-13C NOESY-HSQC ALIPHATIC	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
11	3D 1H-13C NOESY-HSQC AROMATIC	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
12	3D 1H-15N NOESY-HSQC	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800
13	H(C)CONH	1.5 mM [U-13C; U-15N] DolP, 50 mM NA sodium phosphate, 0.02 % w/v NA NaN3, 50 mM NA sodium chloride	90% H2O/10% D2O	0.05 M	6	AMBIENT bar	298	Varian INOVA 800

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	800

NMR Refinement
Method	Details	Software
simulated annealing		ARIA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	THE STRUCTURE WAS CALCULATED USING TRADITIONAL NOE AND DEHEDRAL ANGLE RESTRAINTS

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	ARIA	2.3	LINGE
11	chemical shift assignment	NMRFAM-SPARKY		Lee W, Tonelli M, Markley JL
13	data analysis	NMRFAM-SPARKY		Lee W, Tonelli M, Markley JL
12	structure calculation	CYANA		Guntert, Mumenthaler and Wuthrich
2	refinement	PROCHECK / PROCHECK-NMR		Laskowski, MacArthur, Smith, Jones, Hutchinson, Morris, Moss and Thornton
3	refinement	CYANA	2.1	LASKOWSKI AND MACARTHUR (PROCHECKNMR)
5	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
7	data analysis	TALOS		Cornilescu, Delaglio and Bax

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.