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长春花中海枣苷空泡输入蛋白的特性。

Characterization of a vacuolar importer of secologanin in Catharanthus roseus.

机构信息

Plant Science, McGill University, Sainte-Anne-de-Bellevue, QC, H9X 3V9, Canada.

Chemistry, University of New Brunswick, Fredericton, NB, E3B 5A3, Canada.

出版信息

Commun Biol. 2024 Aug 3;7(1):939. doi: 10.1038/s42003-024-06624-5.

DOI:10.1038/s42003-024-06624-5
PMID:39097635
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11298008/
Abstract

Monoterpenoid indole alkaloid (MIA) biosynthesis in Catharanthus roseus is a paragon of the spatiotemporal complexity achievable by plant specialized metabolism. Spanning a range of tissues, four cell types, and five cellular organelles, MIA metabolism is intricately regulated and organized. This high degree of metabolic differentiation requires inter-cellular and organellar transport, which remains understudied. Here, we have characterized a vacuolar importer of secologanin belonging to the multidrug and toxic compound extrusion (MATE) family, named CrMATE1. Phylogenetic analyses of MATEs suggested a role in alkaloid transport for CrMATE1, and in planta silencing in two varieties of C. roseus resulted in a shift in the secoiridoid and MIA profiles. Subcellular localization of CrMATE1 confirmed tonoplast localization. Biochemical characterization was conducted using the Xenopus laevis oocyte expression system to determine substrate range, directionality, and rate. We can confirm that CrMATE1 is a vacuolar importer of secologanin, translocating 1 mM of substrate within 25 min. The transporter displayed strict directionality and specificity for secologanin and did not accept other secoiridoid substrates. The unique substrate-specific activity of CrMATE1 showcases the utility of transporters as gatekeepers of pathway flux, mediating the balance between a defense arsenal and cellular homeostasis.

摘要

长春花中的单萜吲哚生物碱（MIA）生物合成是植物特化代谢可实现时空复杂性的典范。MIA 代谢跨越一系列组织、四种细胞类型和五个细胞器官，受到精细的调控和组织。这种高度的代谢分化需要细胞间和细胞器间的运输，但这方面的研究还很不足。在这里，我们鉴定了长春花中的一个属于多药和毒性化合物外排（MATE）家族的 secologanin 液泡输入蛋白，命名为 CrMATE1。MATE 的系统发育分析表明 CrMATE1 可能参与生物碱的运输，在两种长春花品种中的体内沉默导致了 secoiridoid 和 MIA 谱的变化。CrMATE1 的亚细胞定位证实了液泡膜的定位。使用非洲爪蟾卵母细胞表达系统进行了生化特性分析，以确定底物范围、方向性和速率。我们可以确认 CrMATE1 是 secologanin 的液泡输入蛋白，在 25 分钟内转运 1mM 的底物。该转运蛋白对 secologanin 具有严格的方向性和特异性，不接受其他 secoiridoid 底物。CrMATE1 的独特底物特异性活性展示了转运蛋白作为途径通量的守门员的效用，介导了防御武器库和细胞内稳态之间的平衡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/044f8b553987/42003_2024_6624_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/20038f7a73d8/42003_2024_6624_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/eac864d952f3/42003_2024_6624_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/0ea0517bb08b/42003_2024_6624_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/01603ae545b3/42003_2024_6624_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/e305ff4de20d/42003_2024_6624_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/044f8b553987/42003_2024_6624_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/20038f7a73d8/42003_2024_6624_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/eac864d952f3/42003_2024_6624_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/0ea0517bb08b/42003_2024_6624_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/01603ae545b3/42003_2024_6624_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/e305ff4de20d/42003_2024_6624_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91f5/11298008/044f8b553987/42003_2024_6624_Fig6_HTML.jpg

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