Immunoelectron Microscopic Observation of the Behaviors of Peroxisomal Enzymes Inducibly Synthesized in an n-Alkane-Utilizable Yeast Cell, Candida tropicalis

Cell Structure and Function

Naomi Kamasawa¹  Itsuko Ohtsuka¹  Atsuo Tanaka²  Mitsuyoshi Ueda²  Yasushi Kamada²  Masako Osumi¹ 

[1] Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University;Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University

关键词: immunoelectron microscopy;    yeast;    Candida tropicalis;    peroxisome (microbody);    β-oxidation enzymes;   

DOI  :  10.1247/csf.21.117

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(16)Cited-By(10)We reported that immunoelectron microscopy was an excellent tool for determining the subcellular localization of thiolase isozymes, acetoacetyl-CoA thiolase (T-I) and 3-ketoacyl-CoA thiolase (T-III) in n-alkane-grown Candida tropicalis cells (KAMASAWA, N. et al., (1992). Cell Struct. Funct., 17: 203-207). Current investigation on the visualization of other peroxisomal enzymes, acyl-CoA oxidase (ACO), catalase (KAT), carnitine acetyltransferase (CAT), isocitrate lyase (ICL) and malate synthase (MS), showed that ACO localized in peroxisomes, KAT in peroxisomes and cytoplasm, and CAT in peroxisomes, mitochondria and cytoplasm. Most of ICL and MS were found in peroxisomes. These results agreed with previous biochemical studies and supported the presumed roles of these enzymes. The same technique was applied to study the process of synthesis and localization of these enzymes early in the cultivation period in n-alkane medium when peroxisomes began to proliferate. ACO and T-III were rapidly induced after transfer of cells from glucose- to n-alkanemedia. There was a drastic change of their location from cytoplasm to peroxisomes between 1 h and 2 h after the transfer, while T-I, KAT and CAT were moderately induced in cytoplasm and their location was gradually changed to each organelle. ICL and MS, the key enzymes in the glyoxylate cycle, were already localized in peroxisomes in the glucose-grown cells and respective inducible enzymes also were gradually localized there. This visual analysis is useful for the vivid elucidation of the process of peroxisome proliferation and enzyme transport within a cell.

【 授权许可】

Unknown