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

[摘要]

Clonal BHK cells permanently transfected with the metabotropic glutamate receptor 1α (mGluR1α), 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.

[学科分类] 生物化学/生物物理

[关键词] Metabotropic glutamate receptor;Phosphorylation;Protein kinase C;Desensitization;Inositol trisphosphate;Diacylglycerol;mGluR;metabotropic glutamate receptor;BHK;baby hamster kidney;PLC;phospholipase C;IP;inositol phospholipid;DAG;diacylglycerol;IP3;inositol trisphosphate;PKC;protein kinase C;PKA;protein kinase A;β-ARK;β-adrenergic receptor kinase;CHO;chinese hamster ovary;DCI;dichloroisocoumarin;PdBu;phorbol-12;13-dibutyrate;DMEM;Dulbecco's Modified Eagle Medium;FCS;foetal calf serum;DTT;dithiothreitol;SDS;sodium dodecyl sulphate;PAGE;polyacrylamide gel electrophoresis