Figure 2.

Binding of serum antibodies to ZnT8·GFP in its native conformation. A, analytical size exclusion HPLC revealed a single monodispersed ZnT8·GFP species in detergent micelles (green trace). Adding a proteoliposome-immunized serum to ZnT8·GFP formed ZnT8A-ZnT8·GFP complexes (red trace). Adding a liposome-immunized serum did not affect the ZnT8·GFP chromatogram profile (blue trace). B, formation of a 1:1 stoichiometric ZnT8A-ZnT8·GFP complex at a reduced serum-to-ZnT8·GFP ratio. The complex was only observed with proteoliposome-immunized sera as indicated. C, immunoblotting analysis of the proteoliposome or liposome peak fraction from B. ZnT8·GFP and ZnT8A were detected using an anti-GFP and anti-mouse IgG antibody as indicated.