植物 FYVE 结构域蛋白在内涵体运输中的功能分析。

Functional Analysis of Plant FYVE Domain Proteins in Endosomal Trafficking.

机构信息

Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou, China.

出版信息

Methods Mol Biol. 2020;2177:83-94. doi: 10.1007/978-1-0716-0767-1_8.

DOI:10.1007/978-1-0716-0767-1_8

PMID:32632807

Abstract

The FYVE domain is a double zinc finger-like domain that predominantly binds phosphatidylinositol 3-phosphate. The FYVE domain is usually found in proteins primarily involved in regulating various aspects of endomembrane homeostasis, including endosome tethering, endocytic recycling, membrane protein sorting, and autophagosome maturation. Whereas FYVE domain proteins have been extensively studied in mammals and yeast, only a few FYVE domain proteins have been identified and characterized in plants. Here, by using as an example FREE1 (FYVE DOMAIN PROTEIN REQUIRED FOR ENDOSOMAL SORTING 1), a protein previously identified by us as a critical factor for endosomal trafficking, we describe methods to determine its lipid binding properties and endosomal localization. In addition, we also demonstrate a method to quickly test whether an FYVE domain protein is involved in endosomal sorting in plant cells.

摘要

FYVE 结构域是一种双锌指样结构域，主要结合磷脂酰肌醇 3-磷酸。FYVE 结构域通常存在于主要参与调节内质网稳态的各种方面的蛋白质中，包括内体连接、内吞体再循环、膜蛋白分选和自噬体成熟。虽然 FYVE 结构域蛋白在哺乳动物和酵母中得到了广泛研究，但在植物中只鉴定和表征了少数 FYVE 结构域蛋白。在这里，我们以 FREE1（内体分选所需的 FYVE 结构域蛋白 1）为例，这是我们之前鉴定的参与内体运输的关键因子，描述了确定其脂质结合特性和内体定位的方法。此外，我们还展示了一种快速测试 FYVE 结构域蛋白是否参与植物细胞内体分选的方法。