Figure 2.
Figure 2. DNA interacts with a region of positive charges at the RAGE dimer interface. (A) Binding in solution of biotinylated ODN 2006 (CpG-B) to His-tagged full length human RAGE extracellular domain (V-C1-C2-His) or isolated structural domains was evaluated by AlphaScreen homogenous binding assay. Indicated concentrations of CpG-B were incubated with 40 nM RAGE. Data shown are from one experiment representative of three independent experiments. (B) Electrostatic charge surface of RAGE is shown on a scale of −10 kT/e (red) to +10 kT/e (blue) in three different orientations. The bound dsDNA is shown as an orange ribbon. (C) Polyethyleneimine (PEI) was incubated together with RAGE and biotinylated CpG DNA, as indicated, and RAGE–DNA interaction was assessed by AlphaScreen assay (PO, phosphodiester and PS, phosphorothioate backbones), CpG-A, and CpG-C. Data shown are from one experiment representative of two independent experiments.

DNA interacts with a region of positive charges at the RAGE dimer interface. (A) Binding in solution of biotinylated ODN 2006 (CpG-B) to His-tagged full length human RAGE extracellular domain (V-C1-C2-His) or isolated structural domains was evaluated by AlphaScreen homogenous binding assay. Indicated concentrations of CpG-B were incubated with 40 nM RAGE. Data shown are from one experiment representative of three independent experiments. (B) Electrostatic charge surface of RAGE is shown on a scale of −10 kT/e (red) to +10 kT/e (blue) in three different orientations. The bound dsDNA is shown as an orange ribbon. (C) Polyethyleneimine (PEI) was incubated together with RAGE and biotinylated CpG DNA, as indicated, and RAGE–DNA interaction was assessed by AlphaScreen assay (PO, phosphodiester and PS, phosphorothioate backbones), CpG-A, and CpG-C. Data shown are from one experiment representative of two independent experiments.

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