拟南芥中 PI3P 的亚细胞定位。

Subcellular Localization of PI3P in Arabidopsis.

机构信息

Laboratory of Cell and Molecular Biology and Department of Botany, University of Wisconsin-Madison, Madison, WI, USA.

Department of Biological Sciences, Pusan National University, Busan, Republic of Korea.

出版信息

Methods Mol Biol. 2020;2177:109-118. doi: 10.1007/978-1-0716-0767-1_10.

DOI:10.1007/978-1-0716-0767-1_10

PMID:32632809

Abstract

Phosphatidylinositol-3-phosphate (PI3P) is a signaling phospholipid enriched in the membranes of late endosomes (LE) and vacuoles. PI3P mediates vacuolar and endosomal trafficking through recruiting PI3P-binding effector proteins to the LE. PI3P is produced from phosphatidylinositol by the PI 3-kinase complex containing VACUOLAR PROTEIN SORTING 34 (VPS34). The role of PI3P has been elucidated by using genetically encoded PI3P biosensors. We previously showed that Arabidopsis VPS38, a component of the VPS34 complex, localized at the LE and that VPS38 is essential for proper PI3P distribution in endosomal and vacuolar trafficking routes. In this chapter, we describe methods for microscopic imaging of PI3P using the PI3P biosensor citrine-2 × FYVE and the PI 3-kinase inhibitors.

摘要

磷脂酰肌醇-3-磷酸（PI3P）是一种富含晚期内体（LE）和液泡膜的信号磷脂。PI3P 通过将 PI3P 结合效应蛋白募集到 LE 来介导液泡和内体运输。PI3P 由 PI3-激酶复合物从磷脂酰肌醇产生，该复合物包含液泡蛋白分选 34（VPS34）。通过使用遗传编码的 PI3P 生物传感器，已经阐明了 PI3P 的作用。我们之前表明，拟南芥 VPS38 是 VPS34 复合物的一个组成部分，定位于 LE，并且 VPS38 对于正确分配内体和液泡运输途径中的 PI3P 是必需的。在本章中，我们描述了使用 PI3P 生物传感器 citrine-2 × FYVE 和 PI3-激酶抑制剂进行 PI3P 显微成像的方法。