Okumoto Kanji, Honsho Masanori, Liu Yuqiong, Fujiki Yukio
Department of Biology, Faculty of Sciences, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka, 819-0395, Japan.
Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.
Methods Mol Biol. 2017;1595:213-219. doi: 10.1007/978-1-4939-6937-1_20.
Peroxisomes are essential intracellular organelles that catalyze a number of essential metabolic pathways including β-oxidation of very long chain fatty acids, synthesis of plasmalogen, bile acids, and generation and degradation of hydrogen peroxide. These peroxisomal functions are accomplished by strictly and spatiotemporally regulated compartmentalization of the enzymes catalyzing these reactions. Defects in peroxisomal protein import result in inherited peroxisome biogenesis disorders in humans. Peroxisomal matrix and membrane proteins are synthesized on free ribosomes and transported to peroxisomes in a manner dependent on their specific targeting signals and their receptors. Peroxisomal protein import can be analyzed using a semi-intact assay system, in which targeting efficiency is readily monitored by immunofluorescence microscopy. Furthermore, cytosolic factors required for peroxisomal protein import can be manipulated, suggesting that the semi-intact system is a useful and convenient system to uncover the molecular mechanisms of peroxisomal protein import.
过氧化物酶体是重要的细胞内细胞器，催化许多重要的代谢途径，包括极长链脂肪酸的β-氧化、缩醛磷脂的合成、胆汁酸的合成以及过氧化氢的生成和降解。这些过氧化物酶体功能是通过严格且时空调节的催化这些反应的酶的区室化来实现的。过氧化物酶体蛋白输入缺陷会导致人类遗传性过氧化物酶体生物发生障碍。过氧化物酶体基质蛋白和膜蛋白在游离核糖体上合成，并以依赖于其特定靶向信号及其受体的方式转运至过氧化物酶体。过氧化物酶体蛋白输入可使用半完整分析系统进行分析，其中靶向效率可通过免疫荧光显微镜轻松监测。此外，过氧化物酶体蛋白输入所需的胞质因子可以被操纵，这表明半完整系统是揭示过氧化物酶体蛋白输入分子机制的有用且便捷的系统。
