Mechanisms and Regulation of Rhomboid Intramembrane Proteolysis

菱形膜内蛋白水解的机制和调控

• 批准号: RGPIN-2015-04645
• 负责人: Munter, Lisa
• 金额: $ 2.55万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=708512
• 关键词: Mechanisms; Regulation; Rhomboid; Intramembrane; Proteolysis

During evolution, species acquired new genes through gene duplication as well as through modifications. Obviously, there are substantial evolutionary differences between kingdoms of life such as bacteria, archaea, fungi, plantae and animals. Despite these diversities, it is remarkable that some proteins were conserved in all kingdoms throughout evolution and can be found in mammals as well as in the common bacteria Escherichia coli. Such conservation of proteins implies that they carry fundamental functions, i.e., they are required to ensure normal functioning of the organism throughout all types of life. Such conserved proteins comprises the rhomboid proteases, which we are investigating. Rhomboid proteases are bound in cell membranes and control the release of signaling molecules, which are often essential for the communication between cells. The bacterial rhomboid proteases are well studied and we even know the exact structure of bacterial rhomboid proteins. But despite their conservation up to animals, there are only a few publications describing first studies of their potential functions in mammals. We want to advance the knowledge on mammalian rhomboid proteases since we are convinced that they carry essential biological functions in mammals. To achieve our aims, we follow three research projects: 1) To unravel the function of a rhomboid protease, it is helpful to know the substrate that is processed by the protease. We have recently identified a novel substrate for one rhomboid protease, rhomboid-like-4. With this novel substrate, we will be able to investigate how the enzyme recognizes the substrate and how rhomboid enzymatic activity is regulated. 2) Rhomboid-like-4 and its new substrate both occur in kidney and lung. We hypothesize that rhomboid-like-4 controls the levels of substrates at the cell surface and therefore impacts on the cell-cell communication in kidney and/or lung. We will further investigate with which proteins this rhomboid protease interacts inside the cell. 3) Our third project investigates rhomboid protease-1, a rhomboid about which very little knowledge is available. We are the first research group to experimentally analyze this rhomboid protease: We determined that rhomboid-like-1 occurs in neurons and we are investigating a mouse model with a deletion in this gene, which will tell us about its physiological functions. In summary, the trainees in my laboratory are performing basic biochemical research from the very first steps to investigate this family of remarkable enzymes in mammalian system and to shed light on their physiological functions.

在进化过程中，物种通过基因复制和修改获得新基因。显然，在生命界如细菌、古菌、真菌、植物和动物之间存在着实质性的进化差异。尽管存在这些差异，但值得注意的是，在整个进化过程中，有些蛋白质在所有界中都是保守的，并且可以在哺乳动物和常见的细菌大肠杆菌中找到。蛋白质的这种保守性意味着它们具有基本功能，即，它们被要求确保生物体在所有类型的生命中的正常功能。 这些保守的蛋白质包括菱形蛋白酶，我们正在研究。菱形蛋白酶结合在细胞膜中，并控制信号分子的释放，这些信号分子通常是细胞间通讯所必需的。细菌菱形蛋白酶的研究已经很深入，我们甚至知道细菌菱形蛋白的确切结构。但是，尽管它们在动物中受到保护，但只有少数出版物描述了它们在哺乳动物中的潜在功能的初步研究。我们希望提高对哺乳动物菱形蛋白酶的认识，因为我们相信它们在哺乳动物中具有重要的生物学功能。为了实现我们的目标，我们遵循三个研究项目： 1)为了阐明菱形蛋白酶的功能，了解蛋白酶加工的底物是有帮助的。我们最近发现了一种新的菱形蛋白酶，菱形样-4基板。有了这种新的底物，我们将能够研究酶如何识别底物以及菱形酶活性如何调节。 2)Rhomboid-like-4及其新底物均存在于肾和肺中。我们假设菱形样4控制细胞表面底物的水平，因此影响肾脏和/或肺中的细胞间通讯。我们将进一步研究这种菱形蛋白酶在细胞内与哪些蛋白质相互作用。 3)我们的第三个项目研究菱形蛋白酶-1，一个菱形的知识很少。我们是第一个对这种菱形蛋白酶进行实验分析的研究小组：我们确定了菱形蛋白酶-1发生在神经元中，我们正在研究一种缺失该基因的小鼠模型，这将告诉我们它的生理功能。 总而言之，我实验室的学员从一开始就在进行基础生物化学研究，以研究哺乳动物系统中的这一非凡酶家族，并阐明其生理功能。