title: Luminescent biosensors for real-time monitoring of intracellular cAMP.
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.
authors.affiliation: Promega Corporation, Madison, WI, USA. brock.binkowski@promega.com
authors.forename: Brock F
authors.initials: BF
authors.lastname: Binkowski
authors.forename: Frank
authors.initials: F
authors.lastname: Fan
authors.forename: Keith V
authors.initials: KV
authors.lastname: Wood
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002460:Cell Line; D000242:Cyclic AMP / Q000032:analysis* / Q000378:metabolism; D048888:Fireflies / Q000201:enzymology; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D049408:Luminescent Agents / Q000032:analysis; D008163:Luminescent Measurements / Q000379:methods*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D015398:Signal Transduction; D014162:Transfection
chemical_list: D049408:Luminescent Agents; D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins; D000242:Cyclic AMP; D008156:Luciferases