Imaging of caspase‐3 activation in HeLa cells stimulated with etoposide using a novel fluorescent probe

FEBS Letters

Nagano, Tetsuo²  Tsuruo, Takashi¹  Mashima, Tetsuo¹  Higuchi, Tsunehiko²  Mizukami, Shin²  Kikuchi, Kazuya²  Urano, Yasuteru² 

[1] Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, 1-37-1 Kami-Ikebukuro, Toshima-ku, Tokyo 170-8455, Japan;Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan

关键词: Apoptosis;    Caspase-3;    Fluorescence;    Imaging;    Ac-DEVD-MCA;    acetyl-Asp-Glu-Val-Asp-MCA;    MCA;    4-methylcoumarinyl-7-amide;    Z-Asp-CH2-DCB;    benzyloxycarbonyl-Asp-CH2OC(O)-2;    6-dichlorobenzene;    DMSO;    dimethyl sulfoxide;    GDEVDGVK;    Gly-Asp-Glu-Val-Asp-Gly-Val-Lys;    LY;    lucifer yellow;    CDCF;    6-carboxydichlorofluorescein;    CTMR;    5-carboxytetramethylrhodamine;    CXR;    5-carboxy-X-rhodamine;    HPLC;    high-performance liquid chromatography;    ODS;    octadecylsilica;    PBS;    phosphate-buffered saline;   

DOI  :  10.1016/S0014-5793(99)00755-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Microscopic visualization of intracellular enzyme activity can provide information about the physiological role of the enzyme. Caspases are cysteine proteases that have critical roles in the execution of apoptosis. General fluorometric substrates of caspase-3, such as DEVD-MCA, are unsuitable for imaging because they are excited at short wavelength, so we designed and synthesized novel fluorescent probes that are excited at suitable wavelengths for detecting caspase-3 activity in living cells. Using one of these probes, we succeeded in microscopic visualization of caspase-3-like activity within HeLa cells treated with etoposide. The caspase-3-like activity was increased in the cytosol at first, then expanded to the whole cell.

【 授权许可】

Unknown