Figure 5.

Gβ5 inhibits the interaction between RGS7 and Gαo. His-tagged Gαo (30) was immobilized on the Ni²⁺-NTA beads, and the ³⁵S-labeled RGS7 or its mixture with Gβ5 was applied, batchwise, to the suspension in the presence of 0.1 mM GDP/0.1 mM GDP plus 10 mM NaF and 100 μM AlCl₃ or guanosine 5′-[γ-thio]triphosphate. The beads were washed and eluted with sample buffer for SDS/PAGE. The resin without Gαo was used for control of a nonspecific adsorption. (A) Radioautograms of the fractions from the chromatography resolved by SDS/PAGE followed by radioautography. (B) The amount of the protein in the bands was quantified by the image analysis of the exposed film. The bar graph shows the amount of RGS7 eluted from immobilized (His)₆-Gαo or control Ni²⁺ beads without Gαo. Data were collected from three independent experiments.