Besagni Celine, Piller Lucia Eugeni, Bréhélin Claire
Laboratoire de Physiologie Végétale, Université de Neuchâtel, Neuchâtel, Switzerland.
Methods Mol Biol. 2011;775:223-39. doi: 10.1007/978-1-61779-237-3_12.
Plastoglobules are particles specifically located inside different types of plastids. They mainly contain lipids and proteins and are physically attached to thylakoids. Proteomic studies have underlined the role of plastoglobules in diverse plastid metabolic pathways, such as those producing vitamin K, vitamin E, and carotenoids, and have implicated them in plant response to stress. This chapter describes the isolation of pure and intact plastoglobules from Arabidopsis leaves. The procedure starts with the isolation of intact chloroplasts by centrifugation on a Percoll gradient. Plastoglobules are then separated from the plastid membranes by flotation on a sucrose gradient. Finally, the purity of the plastoglobule fraction is verified by immunoblotting.
质体小球是特定位于不同类型质体内的颗粒。它们主要包含脂质和蛋白质，并在物理上附着于类囊体。蛋白质组学研究强调了质体小球在多种质体代谢途径中的作用，例如那些产生维生素K、维生素E和类胡萝卜素的途径，并表明它们参与植物对胁迫的响应。本章描述了从拟南芥叶片中分离纯净且完整的质体小球的方法。该过程首先通过在Percoll梯度上离心来分离完整的叶绿体。然后通过在蔗糖梯度上漂浮将质体小球与质体膜分离。最后，通过免疫印迹法验证质体小球组分的纯度。
