Imaging of cAMP‐dependent protein kinase activity in living neural cells using a novel fluorescent substrate

FEBS Letters

Omori, Akira²  Higashi, Hideyoshi²  Kudo, Yoshihisa¹  Sato, Kazuki²  Ohtake, Atsuko²  Yoshida, Sachiyo² 

[1] Tokyo University of Pharmacy and Life Science, Horinouchi, Hachioji, Tokyo 192-03, Japan;Mitsubishi Kasei Institute of Life Sciences, Minamiooya, Machida, Tokyo 194, Japan

关键词: cAMP-dependent protein kinase;    Imaging;    Neuron;    Calmodulin;    Glutamate;    Hippocampus;   

DOI  :  10.1016/S0014-5793(97)00970-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In order to visualize the activity of the cAMP-dependent protein kinase (PKA) in living cells, we have constructed a new fluorescence PKA substrate by conjugating a fluorescence probe to a partial amino acid sequence of PKA regulatory domain II which contains a specific autophosphorylation site. The fluorescent peptide was cell-permeable and became phosphorylated when the intracellular cAMP concentration was increased, resulting in a decrease in its fluorescence intensity. In NG108-15 cells, PKA activity was localized to the cytosol around the nucleus. In cultured hippocampal neurons, addition of l-glutamate caused PKA activation associated with increase of the cellular cAMP.

【 授权许可】

Unknown