New approaches to handle old compounds – Studying the ecological function and bioactivity of Microcystins exploiting bio-orthogonal chemistry

处理旧化合物的新方法 â 利用生物正交化学研究微囊藻毒素的生态功能和生物活性

• 批准号: 452935115
• 负责人: Professor Dr. Timo H. J. Niedermeyer
• 金额: --
• 依托单位: Abteilung Pharmazeutische Biologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/452935115?language=en
• 关键词: New; approaches; handle; old; compounds

Microcystins (MCs) are cyclic heptapeptides produced by different cyanobacterial genera, e.g. Microcystis. As major toxins produced during freshwater cyanobacterial blooms, they have a significant impact on the environment, and sometimes even threaten animal or human life. Due to their importance, MCs belong to the best studied cyanobacterial toxins. However, the biological role of MCs for the producing organisms as well as their multifactorial mode of action on eukaryotic cells are still not fully understood. The goal of this project is to contribute to the understanding of the biological functions and the mode of action of these important cyanotoxins. To reach this goal, we will work on answering the following questions: To which proteins bind MCs in MC producing cells of Microcystis? Where are MCs localized in the producing cells? To which proteins bind MCs after uptake into the eukaryotic cell besides the targets known to date?We will address these questions using our newly developed “clickable” MCs. These compounds, produced in situ in Microcystis, feature reactive functional groups that can be used for bio-orthogonal crosslinking techniques, so-called “click reactions“. We can covalently bind these MCs to labels such as fluorescent dyes or gold nanoparticles, and also attach them to polymers such as functionalized agarose beads. The major techniques that will be used in this project are pull-down proteomics to identify to which proteins MCs do bind, and electron microscopy to analyze the localization of MCs in the producing cells. The compounds can also be used for “target fishing” to identify binding partners in eukaryotic cells and for fluorescence microscopy to study MC uptake and distribution in cells. This will enable us to study the biological roles of MCs in greater detail than it has been possible in previous studies.

微囊藻毒素(MC)是由不同的蓝藻属产生的环状七肽，如微囊藻。作为淡水蓝藻水华产生的主要毒素，它们对环境有重大影响，有时甚至威胁到动物或人类的生命。由于其重要性，MCs属于研究得最好的蓝藻毒素。然而，巨噬细胞在生产生物体中的生物学作用以及它们对真核细胞的多因素作用模式仍不完全清楚。该项目的目标是帮助了解这些重要的氰基毒素的生物学功能和作用方式。为了达到这一目标，我们将致力于回答以下问题：微囊藻MC产生细胞中的MC与哪些蛋白结合？MC在生产细胞中定位在哪里？除了已知的靶点外，在摄取到真核细胞后，哪些蛋白质能与MC结合？我们将使用我们新开发的“可点击”MC来解决这些问题。这些化合物在微囊藻中原位产生，具有反应性官能团，可用于生物正交交联技术，即所谓的“点击反应”。我们可以将这些MC共价结合到荧光染料或金纳米颗粒等标签上，也可以将它们连接到聚合物上，如功能化的琼脂糖珠。该项目将使用的主要技术是下拉蛋白质组学，以确定MC确实与哪些蛋白质结合，以及电子显微镜分析MC在产生细胞中的定位。这些化合物还可用于“靶标钓鱼”，以确定真核细胞中的结合伙伴，并用于荧光显微镜研究MC在细胞中的摄取和分布。这将使我们能够比以前的研究更详细地研究MC的生物学作用。