Target Discovery in Ralstonia solanacearum through an Activity-Based Protein Profiling Technique Based on Bioactive Oxadiazole Sulfones
Target Discovery in Ralstonia solanacearum through an Activity-Based Protein Profiling Technique Based on Bioactive Oxadiazole Sulfones
复制标题
通过基于生物活性恶二唑砜的活性蛋白质分析技术发现青枯菌中的靶标
DOI：
10.1021/acs.jafc.9b07192
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发表时间：
2020-02-26
影响因子：
6.1
通讯作者：
Yang, Song
中科院分区：
文献类型：
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作者：
Chen, Biao;Long, Qing-Su;Yang, Song
青枯病是一种极具破坏性和叛逆性的植物病原菌，可引起200多种植物的青枯病。为了探索和发现青枯菌中潜在的靶标，以开发针对这种感染的新的农用化学品，在这里，我们开发了一种典型的基于活性的蛋白质谱技术来发现青枯菌中的靶标，该技术基于生物活性恶二唑砜衍生的基于活性的探针1。通过定量化学蛋白质组学方法，共确定了65个特定靶点，并具有很高的可信度。三种有代表性的蛋白质(甘氨酸裂解系统H蛋白、硫醇过氧化物酶和二氢硫胺S-琥珀酰基转移酶)分别与各自的抗体进行免疫印迹分析。此外，重组硫醇过氧化物酶与探针1的体外相互作用进一步证实了该蛋白是恶二唑砜的靶标。我们期望这些在青枯菌中发现的蛋白质靶标能够刺激针对青枯菌引起的细菌感染的新型农用化学物质的发现和发展。
Ralstonia solanacearum is an extremely destructive and rebellious phytopathogen that can cause bacterial wilt diseases in more than 200 plant species. To explore and discover the potential targets in R. solanacearum for the purpose of developing new agrochemicals targeting this infection, here, we exploited a typical activity-based protein profiling technique for target discovery in R. solanacearum based on an activity-based probe 1 derived from bioactive oxadiazole sulfones. A total of 65 specific targets were identified with high confidence through a quantitative chemical proteomic approach. Three representative proteins (glycine cleavage system H protein, thiol peroxidase, and dihydrolipoamide S-succinyltransferase) were validated as the targets by using the immunoblotting analysis with their respective antibodies. Additionally, the in vitro interaction between the recombinant thiol peroxidase and probe 1 further confirmed that this protein was a target of oxadiazole sulfones. We anticipated that these discovered protein targets in R. solanacearum can stimulate the discovery and development of novel agrochemicals targeting bacterial infections caused by R. solanacearum.