Cytochrome P450 enzymes in the Arabidopsis indole-3-acetonitrile metabolic network: Biological functions and protein-protein interactions

拟南芥吲哚-3-乙腈代谢网络中的细胞色素 P450 酶：生物学功能和蛋白质-蛋白质相互作用

• 批准号: 281071237
• 负责人: Professor Dr. Erich Glawischnig
• 金额: --
• 依托单位: Professur für Mikrobielle Biotechnologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/281071237?language=en
• 关键词: Cytochrome; P450; enzymes; Arabidopsis; indole

As a model for induced chemical defences, the network of indole-3-acetonitrile (IAN) metabolism and camalexin biosynthesis is analysed in Arabidopsis. A set of multiple mutants in biosynthetic genes is generated to evaluate the roles of different subclasses of tryptophan-metabolites in pathogen resistance. Here, the deletion of an entire cluster of 16 cytochrome P450-encoding genes was achieved somatically following a TALE-nuclease-based approach and now stable deletion lines are aimed for. In an untargeted Co-IP approach, physical interaction of the camalexin biosynthetic enzyme CYP71B15 (PAD3) with other P450-enzymes involved in IAN metabolism, as well as with regulatory proteins, was observed. We will now systematically analyse functional protein-protein interactions by targeted Co-IP, Förster resonance energy transfer, yeast-2-hybrid analysis, and functional coexpression in yeast and Nicotiana benthamiana, to gain insight into the formation of metabolons involved in metabolic defence.

作为诱导化学防御的模型，分析了拟南芥中吲哚-3-乙腈（IAN）代谢和camalexin生物合成的网络。产生一组生物合成基因的多个突变体，以评估不同亚类的多糖代谢物在病原体抗性中的作用。在这里，16个细胞色素P450编码基因的整个簇的缺失是在基于TALE-核酸酶的方法之后体细胞地实现的，并且现在稳定的缺失系的目标是。在非靶向Co-IP方法中，观察到了camalexin生物合成酶CYP 71 B15（PAD 3）与IAN代谢中涉及的其他P450酶以及调节蛋白的物理相互作用。我们现在将通过靶向Co-IP，Förster共振能量转移，酵母双杂交分析和酵母和烟草中的功能性共表达系统地分析功能性蛋白质-蛋白质相互作用，以深入了解参与代谢防御的代谢子的形成。

项目成果

The Formation of a Camalexin Biosynthetic Metabolon

骆驼蓬苷生物合成代谢物的形成

• DOI: 10.1105/tpc.19.00403
• 发表时间: 2019-11-01
• 期刊: PLANT CELL
• 影响因子: 11.6
• 作者: Mucha, Stefanie;Heinzlmeir, Stephanie;Glawischnig, Erich
• 通讯作者: Glawischnig, Erich