Adenovirus encoded cyclic nucleotide-gated channels: a new methodology for monitoring cAMP in living cells.
The current, static methodologies for measuring cyclic AMP (cAMP) may underestimate its regulatory properties. Here, we have exploited the Ca2+-conducting properties of cyclic nucleotide-gated (CNG) channels to measure cAMP in live cells, in response to various stimuli. We placed a mutated CNG channel with high sensitivity to cAMP in adenovirus to maximize and render facile its expression in numerous cell types. The ready, continuous nature of the readout contrasted with the traditional approach, which yielded similar static information, but lacked any continuous or interactive qualities. It seems fair to predict that this readily adopted approach will broaden the perception of cAMP signaling.
Fagan
K A
KA
Department of Pharmacology, University of Colorado Health Sciences Center, 4200 East Ninth Ave., Denver, CO 80262, USA.
Schaack
J
J
Zweifach
A
A
Cooper
D M
DM
D000256:Adenoviridae / Q000235:genetics; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D000242:Cyclic AMP / Q000032:analysis*; D054815:Cyclic Nucleotide-Gated Cation Channels; D005822:Genetic Vectors; D007473:Ion Channels / Q000737:chemistry* / Q000235:genetics; D051381:Rats; D015398:Signal Transduction; D014162:Transfection; D014407:Tumor Cells, Cultured
D054815:Cyclic Nucleotide-Gated Cation Channels; D007473:Ion Channels; D000242:Cyclic AMP; D000262:Adenylyl Cyclases