Gheiratmand Ladan, He Cynthia Y
The Lunenfeld-Tanenbaum Research Institute, Toronto, ON, Canada.
Methods Mol Biol. 2015;1270:427-36. doi: 10.1007/978-1-4939-2309-0_29.
The position and function of the Golgi apparatus are tightly coupled with the microtubule-organizing centers (MTOCs) that play important roles in cell growth and polarity. In the unicellular parasite Trypanosoma brucei, the single Golgi apparatus is adjacent to a novel, bilobed structure located at the proximal base of the flagellum, near to the basal body that nucleates the flagellar axoneme. The duplication and segregation of the bilobed structure are tightly coupled to the duplication and segregation of Golgi, ER exit site, basal bodies, and flagellum, suggesting a role of this unique structure in the precise positioning, biogenesis, and inheritance of these single-copied structures during the cell cycle of procyclic T. brucei. Here, we describe in details two different isolation and proteomic methods to characterize the protein components present on or associated with the bilobed structure, which allow further understanding of its function and association with other membranous and cytoskeletal organelles in the parasite.
高尔基体的位置和功能与微管组织中心（MTOC）紧密相连，微管组织中心在细胞生长和极性方面发挥着重要作用。在单细胞寄生虫布氏锥虫中，单个高尔基体毗邻一种位于鞭毛近端基部的新型双叶结构，靠近为鞭毛轴丝提供起始点的基体。双叶结构的复制和分离与高尔基体、内质网出口位点、基体和鞭毛的复制和分离紧密相关，这表明这种独特结构在布氏锥虫前循环期细胞周期中这些单拷贝结构的精确定位、生物发生和遗传过程中发挥着作用。在此，我们详细描述了两种不同的分离和蛋白质组学方法，以表征存在于双叶结构上或与之相关的蛋白质成分，这有助于进一步了解其功能以及与寄生虫中其他膜性和细胞骨架细胞器的关联。
