From the ‡Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Campus UAB, Bellaterra, 08193 Barcelona, Catalonia, Spain.
§Department of Genetics, University of Barcelona, 08028 Barcelona, Catalonia, Spain.
Mol Cell Proteomics. 2018 Jun;17(6):1112-1125. doi: 10.1074/mcp.RA117.000052. Epub 2018 Mar 9.
Activity-based protein profiling (ABPP) is a powerful proteomic technique to display protein activities in a proteome. It is based on the use of small molecular probes that react with the active site of proteins in an activity-dependent manner. We used ABPP to dissect the protein activity changes that occur in the intercellular spaces of tolerant (Hawaii 7996) and susceptible (Marmande) tomato plants in response to , the causing agent of bacterial wilt, one of the most destructive bacterial diseases in plants. The intercellular space -or apoplast- is the first battlefield where the plant faces Here, we explore the possibility that the limited colonization reported in the apoplast of tolerant tomato is partly determined by its active proteome. Our work reveals specific activation of papain-like cysteine proteases (PLCPs) and serine hydrolases (SHs) in the leaf apoplast of the tolerant tomato Hawaii 7996 on infection. The P69 family members P69C and P69F, and an unannotated lipase (Solyc02g077110.2.1), were found to be post-translationally activated. In addition, protein network analysis showed that deeper changes in network topology take place in the susceptible tomato variety, suggesting that the tolerant cultivar might be more prepared to face in its basal state. Altogether this work identifies significant changes in the activity of 4 PLCPs and 27 SHs in the tomato leaf apoplast in response to , most of which are yet to be characterized. Our findings denote the importance of novel proteomic approaches such as ABPP to provide new insights on old and elusive questions regarding the molecular basis of resistance to .
活性蛋白质组学分析(ABPP)是一种强大的蛋白质组学技术,可显示蛋白质组中的蛋白质活性。它基于使用小分子探针,这些探针以依赖于活性的方式与蛋白质的活性部位反应。我们使用 ABPP 来剖析耐受(夏威夷 7996)和易感(马蒙德)番茄植物细胞间隙中发生的蛋白质活性变化,以应对细菌性萎蔫的病原体,这是植物中最具破坏性的细菌性疾病之一。细胞间隙 - 或质外体 - 是植物首先面临的第一个战场。在这里,我们探讨了在耐受番茄的质外体中报道的有限 定殖部分由其活跃的蛋白质组决定的可能性。我们的工作揭示了在 感染后,耐受番茄夏威夷 7996 的叶片质外体中特定的半胱氨酸蛋白酶(PLCP)和丝氨酸水解酶(SH)的激活。发现 P69 家族成员 P69C 和 P69F 以及未注释的脂肪酶(Solyc02g077110.2.1)被翻译后激活。此外,蛋白质网络分析表明,易感番茄品种中网络拓扑结构发生了更深层次的变化,这表明耐受品种在基础状态下可能更有准备面对 。总之,这项工作确定了在番茄叶片质外体中 4 种 PLCP 和 27 种 SH 对 的活性发生了重大变化,其中大多数尚未得到表征。我们的发现表示,新型蛋白质组学方法(如 ABPP)对于提供有关抵抗 分子基础的旧而难以捉摸的问题的新见解非常重要。