Yamashita Shun-Ichi, Oku Masahide, Sakai Yasuyoshi, Fujiki Yukio
Department of Cellular Physiology, Graduate School of Medical and Dental Sciences, Niigata University, 1-757 Asahimachi-dori, Chuo-ku, Niigata, 951-8510, Japan.
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto, 606-8502, Japan.
Methods Mol Biol. 2017;1595:249-255. doi: 10.1007/978-1-4939-6937-1_24.
Peroxisome abundance is tightly regulated according to the physiological contexts, through regulations of both proliferation and degradation of the organelles. Here, we describe detailed methods to analyze processes for autophagic degradation of peroxisomes, termed pexophagy, in yeast organisms. The assay systems include a method for biochemical detection of pexophagy completion, and one for microscopic visualization of specialized membrane structures acting in pexophagy. As a model yeast organism utilized in studies of pexophagy, the methylotrophic yeast Komagataella phaffii (Pichia pastoris) is referred to in this chapter and related information on the studies with baker's yeast (Saccharomyces cerevisiae) is also included. The described techniques facilitate elucidation of molecular machineries for pexophagy and understanding of peroxisome-selective autophagic pathways.
过氧化物酶体的丰度根据生理环境通过细胞器的增殖和降解调节而受到严格调控。在此，我们描述了详细的方法，用于分析酵母生物体中过氧化物酶体自噬降解过程（称为过氧化物酶体自噬）。检测系统包括一种用于生物化学检测过氧化物酶体自噬完成情况的方法，以及一种用于显微镜观察参与过氧化物酶体自噬的特殊膜结构的方法。作为过氧化物酶体自噬研究中使用的模式酵母生物体，本章提及了甲基营养型酵母毕赤酵母（Komagataella phaffii，以前称为巴斯德毕赤酵母），并且还包括了与酿酒酵母研究相关的信息。所描述的技术有助于阐明过氧化物酶体自噬的分子机制，并增进对过氧化物酶体选择性自噬途径的理解。
