Fig. 2.
Mapping the G protein binding site on 5NT. A, schematic diagram of full-length AC5 and GST-tagged NT fragments. B, purified 5NT fragments (2 μM) were incubated with 1 μM G protein subunits (GDP-Gαs or Gβ1γ2) and GST pull-down assay was performed (n = 3). Immunoblots for protein input are shown. C, gel filtration analysis of 5NTΔ144 and Gαs·βγ. Proteins (10 μM) were resolved on tandem Superdex 75/200 columns and analyzed by SDS-PAGE and immunoblotting. Top, 5NTΔ144/Gαs/Gβ1γ2; bottom, 5NTΔ144 alone (n = 2).