Analysis of G protein-mediated activation of phospholipase C in cultured cells

Methods Mol Biol. 2004:237:99-102. doi: 10.1385/1-59259-430-1:99.

Abstract

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.

Publication types

  • Review

MeSH terms

  • Animals
  • COS Cells
  • Enzyme Activation
  • GTP-Binding Proteins / genetics
  • GTP-Binding Proteins / metabolism*
  • Inositol Phosphates / metabolism*
  • Type C Phospholipases / metabolism*

Substances

  • Inositol Phosphates
  • Type C Phospholipases
  • GTP-Binding Proteins