Abstract
To implicate protein kinase C (PKC) or a specific PKC isozyme in the response of a cell to an agonist, one would like to demonstrate activation of PKC in the cell in response to the agonist and to show that specifically blocking this activation blocks the response. However, because of the lipid-dependence of PKC and the paucity of specific PKC activators, inhibitors, and substrates, it is very difficult to assay for the activation of PKC in a cell. In the early years of PKC investigation, it was observed that stimulation of cells with PKC-activating phorbol esters caused a loss of PKC activity from the cytosol (1) and an accompanying gain of activity in the membrane (2). Since then, subcellular translocation of PKC has been observed in response to a variety of stimuli and assay of this translocation can serve as a surrogate for assessment of PKC activation in a cell. Changes in total cellular PKC can be estimated by assay of phosphatidylserine (PS)-dependent PKC activity or of phorbol ester binding capacity in subcellular fractions at varying times after a stimulus; however, some limitations of these assays must be recognized.
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© 2003 Humana Press Inc., Totowa, NJ
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Sando, J.J., Beals, J.K., Hussaini, I.M. (2003). Subcellular Translocation of Protein Kinase C. In: Newton, A.C. (eds) Protein Kinase C Protocols. Methods in Molecular Biology™, vol 233. Humana Press. https://doi.org/10.1385/1-59259-397-6:77
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DOI: https://doi.org/10.1385/1-59259-397-6:77
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