Moving metals: Systematic characterization of metal transporters in Toxoplasma gondii

移动金属：弓形虫金属转运蛋白的系统表征

• 批准号: BB/W014947/1
• 负责人: Clare Harding
• 金额: $ 54.06万
• 依托单位: University of Glasgow
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW014947%2F1
• 关键词: Moving; metals; Systematic; characterization; metal

Cells require metals to create energy and to grow and divide. However, metals cannot pass through the membranes that surround our cells, and instead are transported across membranes by specialised proteins called transporters. This is true of the cells in our bodies, as well as the pathogens which infect us. I am interested in a single-celled parasite called Toxoplasma gondii. This parasite can live in any of the different cells or tissues in our body, making it exceptionally good at adapting to its environment. We know that Toxoplasma requires metals, but we don't know how they are transporting these metals from our cells into the parasite. In this project, we will identify parasite transporter proteins, and investigate how they are functioning within Toxoplasma. To do this, we will first investigate some metal transporters that we know about because they are similar to ones studied in other organisms. We will find out what happens when we remove these transporters, and where they are localised on the cell. But we predict there are types of transporter proteins that are not similar to those we know about. To find these is more challenging. We will make a collection of parasites with every transporter (about 200) deleted. We will grow this collection of parasites in different conditions (e.g. with different levels of metals). By measuring how much they grow, we can find transporters that, when we delete them, change the parasite's ability to grow under different metal conditions. This will help us identify which transporters are important to the parasite in different environments.To investigate how these transporter proteins function, we will also take other approaches. We will see if our parasite transporters can replace ones that are well-studied in yeast. If they can it means that they are likely to have the same or a similar function. We will also put our transporters into frog eggs. Frog eggs are special because they do not have any transporters of their own, so using these large cells we can measure very precisely how our parasite transporters are working to move metals into and out of the cell.In this project, we will learn the identity of Toxoplasma metal transporters and how they function in these parasites. Because metals are essential to the parasite, if we understand this we will understand more about how the parasite produces energy and how it interacts with the cells or our body. This knowledge will help us in the future discover new way to treat Toxoplasma, and possibly its related parasite Plasmodium, which causes malaria.

细胞需要金属来产生能量并生长和分裂。然而，金属不能穿过细胞周围的膜，而是通过称为转运蛋白的特殊蛋白质跨膜转运。我们体内的细胞以及感染我们的病原体都是如此。我对一种叫做弓形虫的单细胞寄生虫感兴趣。这种寄生虫可以生活在我们体内的任何不同细胞或组织中，使其非常善于适应环境。我们知道弓形虫需要金属，但我们不知道它们如何将这些金属从我们的细胞运输到寄生虫中。在这个项目中，我们将鉴定寄生虫转运蛋白，并研究它们如何在弓形虫内发挥作用。为此，我们将首先研究一些我们所知道的金属转运蛋白，因为它们与在其他生物体中研究的金属转运蛋白相似。我们将了解当我们去除这些转运蛋白时会发生什么，以及它们在细胞上的定位。但我们预测有些类型的转运蛋白与我们所知的不相似。找到这些更具挑战性。我们将删除每个传输器（大约 200 个）来收集寄生虫。我们将在不同条件下（例如，使用不同水平的金属）培养这些寄生虫。通过测量它们的生长程度，我们可以找到转运蛋白，当我们删除它们时，会改变寄生虫在不同金属条件下生长的能力。这将帮助我们确定哪些转运蛋白对不同环境中的寄生虫很重要。为了研究这些转运蛋白如何发挥作用，我们还将采取其他方法。我们将看看我们的寄生虫转运蛋白是否可以取代酵母中经过充分研究的转运蛋白。如果可以，则意味着它们很可能具有相同或相似的功能。我们还将把我们的运输机放入青蛙蛋中。青蛙卵很特殊，因为它们没有自己的任何转运蛋白，因此使用这些大细胞，我们可以非常精确地测量寄生虫转运蛋白如何将金属移入和移出细​​胞。在这个项目中，我们将了解青蛙卵的身份弓形虫金属转运蛋白及其在这些寄生虫中的功能。因为金属对于寄生虫至关重要，如果我们了解这一点，我们就会更多地了解寄生虫如何产生能量以及它如何与细胞或我们的身体相互作用。这些知识将帮助我们在未来发现治疗弓形虫以及可能导致疟疾的相关寄生虫疟原虫的新方法。