Mechanisms and functions of protein S-acylation modifications at the plant vacuolar membrane

蛋白质 S-酰化是一种重要的二次修饰，调节蛋白质膜的结合、靶向和功能。然而，修饰蛋白 S-酰基转移酶 (PAT) 和修饰靶蛋白的详细生化和遗传分析因修饰酶针对靶标的功能冗余而受到阻碍。在植物中，蛋白质 S-酰化的机制、功能和要求尚不清楚。因此，我们采用功能筛选并从拟南芥中鉴定出特定的酶-底物对 (PAT10-CBL2)，现在可以直接解决植物液泡膜的脂质修饰机制。因此，该项目旨在阐明来自拟南芥的 PAT 蛋白 10 和 11（代表液泡膜定位酶）的功能特性。此外，我们将分析酶 (PAT) 及其底物靶向所需的机制原理，特别是液泡 PAT 酶对液泡膜结合钙调磷酸酶 B 样 (CBL) 蛋白等底物的特异性修饰。

Protein S-acylation is an important secondary modification and regulates protein membrane association, targeting and function. However, detailed biochemical and genetic analyses of the modifying protein S-acyltransferases (PATs) and of modified target proteins have been hampered by the functional redundancy of modifying enzymes towards targets. In plants, the mechanisms, functions and requirements for protein S-acylation are not well understood. Therefore, we employed a functional screen and identified a specific enzyme-substrate pair (PAT10-CBL2) from Arabidopsis thaliana, which now allows addressing the mechanism of lipid modification at the plant tonoplast directly. Consequently, this project aims to elucidate the functional properties of the PAT proteins 10 and 11 from Arabidopsis, representing tonoplast localized enzymes. In addition we will analyze the mechanistic principles required for the targeting of enzymes (PAT) as well as their substrates, and especially the specific modification of substrates like the tonoplast bound calcineurin B-like (CBL) proteins by the vacuolar PAT enzymes.