Adenovirus encoded cyclic nucleotide‐gated channels: a new methodology for monitoring cAMP in living cells

FEBS Letters

Fagan, Kent A.³  Cooper, Dermot M.F.³  Schaack, Jerome²  Zweifach, Adam¹ 

[1] Department of Physiology, University of Colorado Health Sciences Center, Denver, CO 80262, USA;Department of Microbiology, University of Colorado Health Sciences Center, Denver, CO 80262, USA;Department of Pharmacology, P.O. Box C-236, University of Colorado Health Sciences Center, 4200 E. Nineth Ave., Denver, CO 80262, USA

关键词: Adenylyl cyclase;    Cyclic AMP;    Cyclic nucleotide-gated channel;    Adenovirus;    [Ca2+]i;    cytosolic calcium concentration;    CNG channel;    cyclic nucleotide-gated channel;    PKA;    cAMP-dependent protein kinase;    GFP;    green fluorescent protein;    ddA;    2′;    5′-dideoxyadenosine;    2MeSATP;    2-methylthioadenosine 5′-triphosphate;   

DOI  :  10.1016/S0014-5793(01)02564-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown