Analysis of G protein-mediated activation of phospholipase C in cultured cells.
Inositide-specific phospholipase C (PLC) beta can be activated by the Gq family of G alpha subunits and by the G beta gamma subunits of heterotrimeric G proteins. PLC catalyzes the hydrolysis of phosphotidylinositol 4,5-bisphosphates (PIP2) to inositol trisphosphate (IP3) and diacylglycerol (DAG). This chapter describes a cell-based assay system to determine the activity of PLC by monitoring the levels of IPs in cultured cells. This assay system can be used to examine the activation of G protein-coupled receptors (GPCRs), coupling of GPCRs to G proteins, and the interactions of G proteins to PLC beta.
D000818:Animals; D019556:COS Cells; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D007295:Inositol Phosphates / Q000378:metabolism*; D010738:Type C Phospholipases / Q000378:metabolism*
D007295:Inositol Phosphates; D010738:Type C Phospholipases; D019204:GTP-Binding Proteins