Figure 5. Both Gβγ and PKCα bind distal segments of α_1C-NT. (A) Binding of [³⁵S]-labeled Gβ₁γ₂ to different GST-fused fragments of the NT. Immobilized GST fusion proteins were incubated with Gβ₁γ₂. After washing, Gβ₁γ₂-bound proteins were eluted and resolved by SDS-PAGE. Gβ₁γ₂ was found to bind the distal third of the NT. Similar data was obtained in six more experiments. (B) Binding of [³⁵S]-labeled PKCα to different GST-fused fragments of the NT. PKCα was found to bind the distal half of the NT. (C) [³⁵S]PKCα binding to segments of the NT was repeated in the presence of 1.5 μg purified Gβ₁γ₂ . It did not alter PKC binding to the NT, suggesting no binding competition for the same site.