Mining Ser hydrolase activities from the Pseudomonas-Arabidopsis interaction

从假单胞菌-拟南芥相互作用中挖掘 Ser 水解酶活性

• 批准号: 195647573
• 负责人: Professor Dr. Renier A.L. van der Hoorn
• 金额: --
• 依托单位: Department of Plant Sciences
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/195647573?language=en
• 关键词: Mining; Ser; hydrolase; activities; Pseudomonas

Plant-pathogen interactions comprise a complex network of antagonistic interactions where protein activities of the opponent are manipulated by post-translational modifications, inhibitors and activators. This functional proteomic information can not be displayed with traditional transcriptomic and proteomic approaches. Activity-based protein profiling (ABPP) is a robust and unique, novel technology that displays the activity layer of the proteome using activity-based probes that react with active site residues in a mechanism-dependent manner. This project proposal aims at identifying differential activities of serine hydrolases (SHs) using trifunctional fluorophosphonate (TriFP) probes. TriFP can label over 600 SHs encoded by the Arabidopsis genome, including numerous SHs involved in immunity such as GDSL lipases, EDS1, PAD4, methylesterases and subtilases. Preliminary experiments revealed several differential SH activities upon infection with Pseudomonas syringae, and the aim is to identify these and additional differential SH activities and investigate their role during compatible and incompatible interactions using biochemistry and reverse genetics.

植物-病原体相互作用包括拮抗相互作用的复杂网络，其中对手的蛋白质活性通过翻译后修饰、抑制剂和激活剂来操纵。传统的转录组学和蛋白质组学方法不能显示这些功能蛋白质组学信息。基于活性的蛋白质谱分析（ABPP）是一种强大而独特的新技术，其使用基于活性的探针以机制依赖的方式与活性位点残基反应来显示蛋白质组的活性层。本项目旨在利用三功能氟膦酸酯（TriFP）探针鉴定丝氨酸水解酶（SH）的差异活性。TriFP可以标记拟南芥基因组编码的600多种SH，包括许多参与免疫的SH，如GDSL脂肪酶、EDS 1、PAD 4、甲基酯酶和枯草杆菌酶。初步的实验揭示了几个差异SH活动感染假单胞菌，其目的是确定这些和其他差异SH活动，并调查他们的作用，在兼容和不兼容的相互作用，使用生物化学和反向遗传学。