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FEBS Lett. 1997 Sep 1;414(1):55-60.

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

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Mitsubishi Kasei Institute of Life Sciences, Machida, Tokyo, Japan.


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.

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