7D3X

Non-specific and specific interactions work cooperatively to promote cytidine deamination catalyzed by APOBEC3A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D HNCO	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	90% H2O/10% D2O	375 mM	7.3	1 atm	293	Bruker AVANCE 900
2	3D HNCA	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	90% H2O/10% D2O	375 mM	7.3	1 atm	293	Bruker AVANCE 900
3	3D HNCACB	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	90% H2O/10% D2O	375 mM	7.3	1 atm	293	Bruker AVANCE 900
4	3D CBCA(CO)NH	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	90% H2O/10% D2O	375 mM	7.3	1 atm	293	Bruker AVANCE 900
5	3D HN(CO)CA	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	90% H2O/10% D2O	375 mM	7.3	1 atm	293	Bruker AVANCE 900
6	3D HCCH-TOCSY	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	100% D2O	375 mM	7.3	1 atm	293	Agilent DD2 800
7	2D 1H-13C HSQC	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	100% D2O	375 mM	7.3	1 atm	293	Agilent DD2 800
8	2D 1H-15N HSQC	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	90% H2O/10% D2O	375 mM	7.3	1 atm	293	Bruker AVANCE 900
9	3D 1H-15N TOCSY	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	90% H2O/10% D2O	375 mM	7.3	1 atm	293	Bruker AVANCE 900
10	3D 1H-15N NOESY	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	90% H2O/10% D2O	375 mM	7.3	1 atm	293	Bruker AVANCE 900
11	3D 1H-13C NOESY aliphatic	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	100% D2O	375 mM	7.3	1 atm	293	Bruker AVANCE 900
12	3D 1H-13C NOESY aromatic	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	100% D2O	375 mM	7.3	1 atm	293	Bruker AVANCE 900
13	2D filter TOCSY	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	100% D2O	375 mM	7.3	1 atm	293	Agilent DD2 800
14	2D filter NOESY	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	100% D2O	375 mM	7.3	1 atm	293	Agilent DD2 800
15	3D 15N-filter-edit NOESY	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	90% H2O/10% D2O	375 mM	7.3	1 atm	293	Agilent DD2 800
16	3D 13C-filter-edit NOESY	25 mM sodium phosphate, 200 mM sodium chloride, 50 uM Zinc chloride, 5 mM DTT, 0.28 mM [U-13C; U-15N] A3A, 0.56 mM DNA	100% D2O	375 mM	7.3	1 atm	293	Agilent DD2 800

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	900
2	Agilent	DD2	800

NMR Refinement
Method	Details	Software
simulated annealing		CNS

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

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	NMRDraw		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	chemical shift assignment	Sparky		Goddard
3	structure calculation	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
4	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read

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.