Production of full-length proteins of the COVID encounter complex for structural analysis and drug discovery

SARS-CoV-2病毒改变了世界人民的生活。该病毒是冠状病毒家族的成员，可以感染体内的一系列细胞，最重要的是呼吸道细胞。这种疾病的全球重要性导致了一项广泛的努力，以开发减轻其影响的新疗法。开发这种疗法的一条途径是找到一种方法来阻止病毒进入宿主的细胞。这一过程是由病毒表面的一种蛋白质(Spike)和人类细胞表面的一种蛋白质(ACE2)介导的。这一过程得到了细胞表面其他蛋白质的帮助，其中包括一种名为B0AT1的蛋白质。如果Spike和ACE2之间的这种相互作用可以被药物阻断，那么感染就可以被抑制。不幸的是，所有这些蛋白质都是膜的一部分；要么是病毒周围的膜，要么是细胞周围的膜。这使得制造这些蛋白质在技术上具有挑战性，这意味着很难进行生产新药所需的研究。在伯明翰大学和牛津大学，我们开发了两个新的系统，使我们能够以稳定的形式制造这些蛋白质。这使我们能够对病毒与人类细胞结合的过程进行前所未有的详细研究。在这个项目中，我们将使用这些方法来生产每一种蛋白质，然后将它们组装成触发细胞病毒感染的结构。我们将使用电子显微镜来研究这种结构，以确定可能成为药物靶点的区域。然后，我们将使用相同的蛋白质样本来开发系统，这些系统可以用来测试各种可能抑制这种复合体形成的药物。总而言之，该项目的成功可能导致针对SARS-CoV-2和其他冠状病毒的新疗法。

The virus SARS-CoV-2 has changed the lives of the world population. The virus is a member of the corona virus family that infects a range of cells in the body, the most important being those of the respiratory tract. The global importance of this disease has led to an extensive effort to develop new therapies that mitigate its effects. One route for developing such therapeutics is to find a way of blocking the entry of the virus into the cells of the host. This process is mediated by a protein on the surface of the virus (Spike) that docks with a protein on the surface of human cells (ACE2). This process is aided by other proteins on the cell surface including one called B0AT1. If this interaction between Spike and ACE2 could be blocked by a drug, then the infection could the inhibited. Unfortunately, all of these proteins a part of a membrane; either the membrane that surrounds the virus or the membrane that surrounds the cell. This makes it technically challenging to make these proteins meaning that is can be difficult to carry out the studies required to produce new drugs.At the Universities of Birmingham and Oxford we have developed 2 novel systems that allow us to make these proteins in a stable form. This enables us to study process of viral binding to human cells in unprecedented detail. In this project we will use these methods to produce each protein and then assemble them to form the structure that triggers viral infection of the cell. We will use Electron Microscopy to study this structure to identify regions that could be targeted by drugs. We will then use the same protein samples to develop systems that could be used to test a wide range of drugs that might inhibit the formation of this complex. Taken together, success in this project could lead to new therapies for SARS-CoV-2 and other corona viruses.