6NFW

Potyvirus viral protein genome linked (VPg) emulates the m7G cap to recruit the eukaryotic translation initiation factor eIF4E

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
7	2D 1H-15N HSQC	450 uM [U-13C; U-15N] PVY-VPg, 50 mM sodium phosphate, 150 mM sodium chloride, 1 mM DTT, 0.02 % v/v sodium azide, 100 uM DSS	93% H2O/7% D2O	150 mM	7.5	1 atm	293	Bruker AVANCE III 600
6	2D 1H-13C HSQC	450 uM [U-13C; U-15N] PVY-VPg, 50 mM sodium phosphate, 150 mM sodium chloride, 1 mM DTT, 0.02 % v/v sodium azide, 100 uM DSS	100% D2O	150 mM	7.5	1 atm	293	Bruker AVANCE III 600
8	2D 1H-13C HSQC	450 uM ILV labeled PVY-VPg, 50 mM sodium phosphate, 150 mM sodium chloride, 1 mM DTT, 0.02 % v/v sodium azide, 100 uM DSS	93% H2O/10% D2O	150 mM	7.5	1 atm	293	Bruker AVANCE III 600
1	3D 1H-15N NOESY	450 uM [U-13C; U-15N] PVY-VPg, 50 mM sodium phosphate, 150 mM sodium chloride, 1 mM DTT, 0.02 % v/v sodium azide, 100 uM DSS	93% H2O/7% D2O	150 mM	7.5	1 atm	293	Bruker AVANCE III 800
2	3D 1H-13C NOESY	450 uM [U-13C; U-15N] PVY-VPg, 50 mM sodium phosphate, 150 mM sodium chloride, 1 mM DTT, 0.02 % v/v sodium azide, 100 uM DSS	93% H2O/7% D2O	150 mM	7.5	1 atm	293	Bruker AVANCE III 800
3	3D 1H-13C NOESY	450 uM [U-13C; U-15N] PVY-VPg, 50 mM sodium phosphate, 150 mM sodium chloride, 1 mM DTT, 0.02 % v/v sodium azide, 100 uM DSS	100% D2O	150 mM	7.5	1 atm	293	Bruker AVANCE III 800
4	4D methyl-methyl 13C-resolved HMQC-NOESY-HMQC	450 uM ILV labeled PVY-VPg, 50 mM sodium phosphate, 150 mM sodium chloride, 1 mM DTT, 0.02 % v/v sodium azide, 100 uM DSS	93% H2O/10% D2O	150 mM	7.5	1 atm	293	Bruker AVANCE III 800
5	3D 1H-13C NOESY aromatic	450 uM [U-13C; U-15N] PVY-VPg, 50 mM sodium phosphate, 150 mM sodium chloride, 1 mM DTT, 0.02 % v/v sodium azide, 100 uM DSS	100% D2O	150 mM	7.5	1 atm	293	Bruker AVANCE III 800

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE III	600
2	Bruker	AVANCE III	800

NMR Refinement
Method	Details	Software
simulated annealing		CYANA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	100
Conformers Submitted Total Number	10
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	CYANA	3.98	Guntert, Mumenthaler and Wuthrich
2	structure calculation	CYANA	3.98	Guntert, Mumenthaler and Wuthrich
3	chemical shift assignment	Sparky		Goddard
4	peak picking	Sparky		Goddard

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.