Rapid agonist mediated phosphorylation of the metabotropic glutamate receptor 1α by protein kinase C in permanently transfected BHK cells

Abstract

Clonal BHK cells permanently transfected with the metabotropic glutamate receptor 1 alpha (mGluR1 alpha), which is coupled to phospholipase C, were used to study the phosphorylation state of the receptor. Cells were labelled with 32PO4(3-), lysed, the receptor immunoprecipitated with specific anti-peptide antibodies and the immunoprecipitates analysed by SDS-PAGE followed by autoradiography. A significant basal level of receptor phosphorylation was observed which was rapidly and transiently increased in response to agonist activation of the receptor. This agonist effect was found to be dose dependent with a rapid time course and could be abolished by the specific PKC inhibitor Ro318220, suggesting that PKC was responsible for the agonist mediated phosphorylation of the receptor.