Three-dimensional analysis of protein aggregate body in Saccharomyces cerevisiae cells

被引:9
作者
Kamasawa, N
Yoshida, T
Ueda, M
Tanaka, A
Osumi, M [1 ]
机构
[1] Japan Womens Univ, Fac Sci, Dept Chem & Biol Sci, Bunkyo Ku, Tokyo 1128681, Japan
[2] Japan Womens Univ, Grad Sch, Div Mat & Biol Funct Sci, Bunkyo Ku, Tokyo 1128681, Japan
[3] Kyoto Univ, Grad Sch Engn, Dept Synthet Chem & Biol Chem, Lab Appl Biol Chem,Sakyo Ku, Kyoto 6068317, Japan
来源
JOURNAL OF ELECTRON MICROSCOPY | 1999年 / 48卷 / 02期
关键词
Saccharomyces cerevisiae; peroxisome; protein aggregate body (PAB); 3D reconstruction; high-pressure freezing; confocal fluorescence microscope;
D O I
10.1093/oxfordjournals.jmicro.a023664
中图分类号
TH742 [显微镜];
学科分类号
摘要
Modified genes of peroxisomal isocitrate lyase of Candida tropicalis (CT-ICL) were constructed and expressed in Saccharomyces cerevisiae cells. We observed subcellular localization of expressed products of the mutant CT-ICL genes by immunoelectron microscopy. An unknown structure termed a protein aggregate body (PAB) storing the expressed product was observed in cytoplasm in various mutants (Kamasawa ct al. (1996) J. Electron Microsc. 45:491-497). We chose two typical cells harbouring the mutant ICL genes Delta 55Q and Delta 237-339 to analyse the ultrastructure and three-dimensional (3D) structure of PABs. The PABs had a homogeneous matrix with a wavy periphery in the cell image using a high-pressure freezing fixation method. Although PABs could not be separated from the cytoplasm or mitochondria under a confocal fluorescence microscope, 3D reconstruction of serial electron micrographs clearly showed the PAB was an independent structure of varying size and had the shape of an incomplete sphere. A cell was sometimes observed to have multiple PABs.
引用
收藏
页码:173 / 176
页数:4
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