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Biological Phenomena, Cell Phenomena, and Immunity
Department of Pathology, Anatomy, and Cell Biology Faculty Papers
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The Use Of Time-Resolved Fluorescence Imaging In The Study Of Protein Kinase C Localisation In Cells., Christopher D Stubbs, Stanley W Botchway, Simon J Slater, Anthony W Parker
The Use Of Time-Resolved Fluorescence Imaging In The Study Of Protein Kinase C Localisation In Cells., Christopher D Stubbs, Stanley W Botchway, Simon J Slater, Anthony W Parker
Department of Pathology, Anatomy, and Cell Biology Faculty Papers
BACKGROUND: Two-photon-excitation fluorescence lifetime imaging (2P-FLIM) was used to investigate the association of protein kinase C alpha (PKCalpha) with caveolin in CHO cells. PKCalpha is found widely in the cytoplasm and nucleus in most cells. Upon activation, as a result of increased intracellular Ca2+ and production of DAG, through G-protein coupled-phospholipase C signalling, PKC translocates to a variety of regions in the cell where it phosphorylates and interacts with many signalling pathways. Due to its wide distribution, discerning a particular interaction from others within the cell is extremely difficult. RESULTS: Fluorescence energy transfer (FRET), between GFP-PKCalpha and DsRed-caveolin, was used …