Activity-based proteomics uncovers suppressed hydrolases and a neo-functionalised antibacterial enzyme at the plant-pathogen interface.
Activity-based proteomics uncovers suppressed hydrolases and a neo-functionalised antibacterial enzyme at the plant-pathogen interface.
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基于活性的蛋白质组学揭示了植物-病原体界面上受抑制的水解酶和新功能化的抗菌酶。
DOI：
10.1111/nph.18857
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Sueldo DJ
中科院分区：
文献类型：
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作者：
Sueldo DJ
植物组织的细胞外空间含有数百种可能伤害殖民微生物的水解酶。成功的病原体可能会抑制这些水解酶，使疾病。本文报道了本氏烟草（Nicotiana benthamianaupon）感染假单胞菌（Pseudomonas benthamiae）后细胞外水解酶的动态变化。利用基于活性的蛋白质组学技术，采用生物素标记的探针，同时监测了171种活性水解酶，包括109种丝氨酸水解酶（SH），49种糖苷酶（GH）和13种半胱氨酸蛋白酶（CP）。其中82种水解酶的活性分别为：丝氨酸蛋白酶（SH），半胱氨酸蛋白酶（CP）和丝氨酸蛋白酶（GH）。在感染过程中，60种水解酶（主要是SH）的活性增加，而60种水解酶（主要是GH和CP）的活性在感染过程中受到抑制。活性β-半乳糖苷酶-1（BGAL 1）是受抑制的水解酶之一，与P产生BGAL 1抑制剂一致。你好另一种被抑制的水解酶，发病机制相关的NbPR 3，在短暂过表达时会降低细菌生长。这取决于它的活性位点，揭示了NbPR 3活性在抗菌免疫中的作用。尽管被注释为几丁质酶，但NbPR 3不具有几丁质酶活性，并且含有对抗菌活性至关重要的E112 Q活性位点取代，并且仅存在于烟草物种中。这项研究引入了一种强有力的方法来揭示细胞外免疫的新组分，例如，发现了新功能化的烟草特异性抗菌剂NbPR 3。
The extracellular space of plant tissues contains hundreds of hydrolases that might harm colonising microbes. Successful pathogens may suppress these hydrolases to enable disease. Here, we report the dynamics of extracellular hydrolases inNicotiana benthamianaupon infection withPseudomonas syringae.Using activity‐based proteomics with a cocktail of biotinylated probes, we simultaneously monitored 171 active hydrolases, including 109 serine hydrolases (SHs), 49 glycosidases (GHs) and 13 cysteine proteases (CPs).The activity of 82 of these hydrolases (mostly SHs) increases during infection, while the activity of 60 hydrolases (mostly GHs and CPs) is suppressed during infection. Active β‐galactosidase‐1 (BGAL1) is amongst the suppressed hydrolases, consistent with production of the BGAL1 inhibitor byP. syringae. One of the other suppressed hydrolases, the pathogenesis‐relatedNbPR3, decreases bacterial growth when transiently overexpressed. This is dependent on its active site, revealing a role forNbPR3 activity in antibacterial immunity.Despite being annotated as a chitinase,NbPR3 does not possess chitinase activity and contains an E112Q active site substitution that is essential for antibacterial activity and is present only inNicotianaspecies.This study introduces a powerful approach to reveal novel components of extracellular immunity, exemplified by the discovery of the suppression ofneo‐functionalisedNicotiana‐specific antibacterialNbPR3.