Elucidation of the Rotavirus NSP4-Caveolin-1 and -Cholesterol Interactions Using Synthetic Peptides
Figure 1
Direct Fluorescence-Binding Assays of NSP4112-140 with Caveolin-1 Peptides. (a) Fluorescence spectra of Cy3-C-Cav161-178 titrated with increasing concentrations of NSP4112-140. Spectrum 1 : 100 nM Cy3-C-Cav161-178 in buffer only. Spectra 2–11 : 100 nM Cy3-C-Cav161-178 in the presence of 10, 100, 150, 200, 250, 300, 350, 400, 450, and 500 nM of NSP4112-140, respectively. (b) Fluorescence spectra of Cy3-C-Cav161-178 titrated with increasing concentrations of NSP4150-175 (negative control). Plots (c)–(e) show maximal fluorescence emission (measured at 565 nm upon excitation at 550 nm) for NSP4112-140 in the presence of 25 nM Cy3-N-Cav2-20 (c), 50 nM Cy3-N- Cav19-40 (d), and 100 nM Cy3-C-Cav161-178. (e). Insets: linear plots of the binding curve of NSP4112-140.
