In situ observation of mobility and anchoring of PKCβI in plasma membrane

FEBS Letters

Tamura, Mamoru²  Saito, Kenta²  Ito, Eiko¹  Kinjo, Masataka²  Takakuwa, Yuichi¹ 

[1] Department of Biochemistry, School of Medicine, Tokyo Women's Medical College, Shinjuku, Tokyo 162-8666, Japan;Laboratory of Supramolecular Biophysics, Research Institute for Electronic Science, Hokkaido University, N12W6, Kita-ku, Sapporo 060-0812, Japan

关键词: Fluorescence correlation spectroscopy;    Protein kinase C;    Green fluorescent protein;    Diffusion constant;    Signal transduction;    Plasma membrane;    FCS;    fluorescence correlation spectroscopy;    FAF;    fluorescence autocorrelation function;    EGFP;    enhanced green fluorescent protein;    PKC;    protein kinase C;    DMEM;    Dulbecco's modified Eagle's medium;    HBSS;    Hanks’ balanced salt solution;    LSM;    laser scanning microscope;    Rh6G;    rhodamine 6G;    TPA;    12-O-tetradecanoylphorbol-13-acetate;   

DOI  :  10.1016/S0014-5793(03)00324-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 PKCβI labeled with enhanced green fluorescent protein freely diffusing in cytosol. Upon 12-O-tetradecanoylphorbol-13-acetate activation, a large part of PKCβI is anchored in the plasma membrane but some PKCβI still moves freely near the plasma membrane. These results indicate that a diffusion-driven transport mechanism is appropriate for the molecular mechanism of the PKCβI localization change.

【 授权许可】

Unknown