Figure 2.
Validation of p7 NMR structure in methanol using specific drug interactions. (A) Isotopically labeled wild-type or Leu20Phe FLAG-p7 was tested for rimantadine sensitivity in dye release assays. Black lines, protein only; red line, protein plus 40 µM rimantadine; L, liposomes only; S, solvent control. (B) 1H/15N HSQC spectra of wild-type and Leu20Phe p7 in the presence (red) or absence (black) of a 2 molar excess of rimantadine. Specific minimal chemical shift changes in the presence/absence of drug were detectable in wild-type, but not Leu20Phe resistant protein (see Supporting Fig. 2c for chemical shift change in parts per million (Δppm) values). (C) Positions of residues on monomeric structure seen to experience chemical shift changes in the presence of rimantadine, including Leu20. Graded color represents magnitude of Δppm; pink (0.14-0.25), red (0.25-0.5), dark red (0.5+).