title: In situ observation of mobility and anchoring of PKCbetaI in plasma membrane.
abstract: We employed fluorescence correlation spectroscopy (FCS) to analyze the characteristics of biomolecules in living cells. Protein kinase C (PKC) changes its subcellular localization from cytosol to the plasma membrane by its ligand. Using FCS, we found PKCbetaI labeled with enhanced green fluorescent protein freely diffusing in cytosol. Upon 12-O-tetradecanoylphorbol-13-acetate activation, a large part of PKCbetaI is anchored in the plasma membrane but some PKCbetaI still moves freely near the plasma membrane. These results indicate that a diffusion-driven transport mechanism is appropriate for the molecular mechanism of the PKCbetaI localization change.
authors.affiliation: Laboratory of Supramolecular Biophysics, Research Institute for Electronic Science, Hokkaido University, Sapporo, Japan.
authors.forename: Kenta
authors.initials: K
authors.lastname: Saito
authors.forename: Eiko
authors.initials: E
authors.lastname: Ito
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Takakuwa
authors.forename: Mamoru
authors.initials: M
authors.lastname: Tamura
authors.forename: Masataka
authors.initials: M
authors.lastname: Kinjo
mesh_terms: D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology*; D004058:Diffusion; D049452:Green Fluorescent Proteins; D006801:Humans; D007700:Kinetics; D008164:Luminescent Proteins / Q000235:genetics; D011493:Protein Kinase C / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D064546:Protein Kinase C beta; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000032:analysis; D013050:Spectrometry, Fluorescence; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D013755:Tetradecanoylphorbol Acetate