Figure 3.
GABAAR phosphorylation by intimately associated PKC is decreased in SE. A, B, Phosphorylation of the β3 by intimately associated PKC activity. A, Top, Detergent-solubilized hippocampal extracts were immunoprecipitated with IgG or anti-β3 antibody, and precipitated material was incubated with 10 μCi 32P-γATP at 30°C for 15 min followed by SDS-PAGE. A, Bottom, The level of β3 subunit phosphorylation was then determined for control and SE extracts, and the level of β3 subunit phosphorylation was normalized to that seen with control extracts (100%). B, Phosphorylation was also assessed in the absence (−) or presence (+) of 15 μm GF 109203. C, The levels of the α, β, and γ PKC isoforms associated with GABAAR are reduced in SE. Material immunoprecipitating with IgG or anti-β3 from control and SE extracts was immunoblotted with an antibody specific to the PKC α, β, and γ isoforms. In, 10% of the input used for each immunoprecipitation.