Untargeted Proteomics Identifies Plant Substrates of the Bacterial-Derived ADP-Ribosyltransferase AvrRpm1.

IF 2.8 3区生物学 Q2 PLANT SCIENCES

Plant Direct Pub Date : 2025-11-16 eCollection Date: 2025-11-01 DOI:10.1002/pld3.70115

Simranjit Kaur, Thomas Colby, Domenika Thieme, Carsten Proksch, Susanne Matschi, Ivan Matić, Lennart Wirthmueller

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引用次数: 0

Abstract

One class of enzymes that plant pathogens employ to manipulate innate immunity and physiology of the infected cells is host-targeted ADP-ribosyltransferases. The bacterial pathogen Pseudomonas syringae uses its Type III secretion system to inject several effector proteins with ADP-ribosyltransferase activity into plant cells. One of them, AvrRpm1, ADP-ribosylates the plasma membrane-associated RPM1-INTERACTING PROTEIN 4 (RIN4) in Glycine max and Arabidopsis thaliana to attenuate targeted secretion of defense-promoting compounds. Substrate identification of host-targeted ADP-ribosyltransferases is complicated by the biochemical lability of the protein modification during plant protein extraction and in several cases requires prior knowledge of plant immune signaling pathways that are impaired by the ADP-ribosylating Type III effector. Using the AvrRpm1-RIN4 pair as a proof of concept, we present an untargeted proteomics workflow for enrichment and detection of ADP-ribosylated proteins and peptides from plant cell extracts that in several cases provides site resolution for the modification.

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非靶向蛋白质组学鉴定细菌来源的adp -核糖基转移酶AvrRpm1的植物底物。

植物病原体利用一类酶来操纵受感染细胞的先天免疫和生理，即宿主靶向adp -核糖基转移酶。细菌病原体丁香假单胞菌利用其III型分泌系统将几种具有adp -核糖基转移酶活性的效应蛋白注入植物细胞。其中一种是AvrRpm1， adp核糖化了甘氨酸和拟南芥中质膜相关的rpm1相互作用蛋白4 (RIN4)，以减弱防御促进化合物的靶向分泌。宿主靶向adp核糖基转移酶的底物鉴定由于植物蛋白提取过程中蛋白质修饰的生化不稳定性而变得复杂，并且在一些情况下需要事先了解被adp核糖基化III型效应物破坏的植物免疫信号通路。利用AvrRpm1-RIN4对作为概念证明，我们提出了一种非靶向蛋白质组学工作流程，用于从植物细胞提取物中富集和检测adp核糖化蛋白和肽，在一些情况下为修饰提供了位点分辨率。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Plant Direct Environmental Science-Ecology

CiteScore

5.00

自引率

3.30%

发文量

101

审稿时长

14 weeks

期刊介绍： Plant Direct is a monthly, sound science journal for the plant sciences that gives prompt and equal consideration to papers reporting work dealing with a variety of subjects. Topics include but are not limited to genetics, biochemistry, development, cell biology, biotic stress, abiotic stress, genomics, phenomics, bioinformatics, physiology, molecular biology, and evolution. A collaborative journal launched by the American Society of Plant Biologists, the Society for Experimental Biology and Wiley, Plant Direct publishes papers submitted directly to the journal as well as those referred from a select group of the societies’ journals.