Figure 1
From: Limited high-throughput screening compatibility of the phenuivirus cap-binding domain
Fluorescence polarization experiments. (a) The figure shows crystal structures of SFTSV CBD (6XYA), PB2 CBD (2VQZ) and eIF4E (1L8B) in complex with m7GTP (presented as lines in black). Proteins are displayed as ribbon diagram with the side chains involved in the m7GTP interaction shown as sticks. The figure was created with PyMOL 2.5. (b) Structural formulas of the employed tracers: m7GTP-γ-aminophenyl-PEG4-Cy5 and EDA-m7GTP-Cy5 (Jena Bioscience GmbH). (c) Detection of the interaction of SFTSV CBD with the different tracers (upper panel: m7GTP-γ-aminophenyl-PEG4-Cy5, lower panel: EDA-m7GTP-Cy5) and (d) of the interaction of PB2 CBD and eIF4E with EDA-m7GTP-Cy5 via FP. The FP signal was measured in presence of 5 nM tracer and the results were fitted according to a single site binding model. Measurements were done in technical triplicates and are shown as representatives of at least 3 biological replicates (all biological replicates are shown in Supplementary Fig. 3). Fitting and plotting was done in GraphPad Prism 9.5.
