Visualisation of intracellular signaling using GFP-fusion protein
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概要
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Protein kinase C (PRO), a family of phospholipiddependent serine/threonine kinases consisting of at least 12 subtypes, plays an important role in various cellular signal tranaductions. It is important to identify the individual function of each PKC subtype for understanding the complicated roles of protein kinase C in various signaling pathways. Studies for the specific subcellular localization of each subtypes has been considered to be one of the most attractive ways to know the individual role of PKC subtypes. Activation of PKC, however, induces its translocation to various cellular compartments, suggesting that PKC targeting to the specific compartment controls the subtype-specific function of PKC in the signaling pathways. Using GFP-tagged fusion protein, we have analyzed the translocation of PKC in living cells and demonstrated the subtype-specific translocation of PKC subtypes. We have demonstrated that the activation of each PKC subtype through different stimuli induces distinct translocations and the different PKC translocation leads to distinct cellular responses. The results showed that subtypespecific and cell-specific PKC targeting is very important for the association with PKC substrate and also for the modulation of cellular functions. Furthermore, we made tranagenic mice expressing GFP-tagged PKC subtypes under the control of various promoter using tetracyclin-regulated system. The translocation of PKC-GFP was studied in living brain slices, indicating that PKC translocation induced by the same stimulation varied in cell types. The results suggested that the each PKC subtype varies its targeting-site in response to each signal and also varies in each cells, and that the targeting to the specific compartment is necessary for the phosphorylation of a specific substrate.
- 日本組織細胞化学会の論文
著者
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Saito Naoaki
Laboratory Of Molecular Pharmacology Biosignal Research Center Kobe University
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Saito Naoaki
Laboratory Of Molecular Pharmacology Biosignal Research Center Kobe Univ.
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