Luminescent biosensors for real-time monitoring of intracellular cAMP

Methods Mol Biol. 2011;756:263-71. doi: 10.1007/978-1-61779-160-4_14.

Abstract

G-protein coupled, seven-transmembrane (7-TM) receptors (GPCRs) comprise a diverse class of signaling molecules involved in cellular physiology and pathology. In recent years, intracellular biosensors have been developed to monitor changes in intracellular cAMP in real time, facilitating studies on the mechanisms of GPCR activation and desensitization in living cells. However, methods based on fluorescence can show limitations in response dynamics together with being difficult to perform. Here we present the use of genetically encoded, luminescent biosensors that allow a facile, non-lytic assay format for monitoring cAMP dynamics in living cells.

MeSH terms

  • Animals
  • Biosensing Techniques / methods*
  • Cell Line
  • Cyclic AMP / analysis*
  • Cyclic AMP / metabolism
  • Fireflies / enzymology
  • Humans
  • Luciferases / analysis
  • Luciferases / genetics
  • Luminescent Agents / analysis
  • Luminescent Measurements / methods*
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism*
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / genetics
  • Signal Transduction
  • Transfection

Substances

  • Luminescent Agents
  • Receptors, G-Protein-Coupled
  • Recombinant Fusion Proteins
  • Cyclic AMP
  • Luciferases