Deciphering the impact of Sars-CoV-2 envelope protein glycosylation on human pathogenicity

正在进行的冠状病毒病2019年(新冠肺炎)大流行是由冠状病毒SARS-CoV-2引起的。病毒进入人类宿主细胞是由与血管紧张素转换酶2(ACE2)结合的糖蛋白Spike S介导的。与其他病毒膜蛋白相比，斯派克S的N-糖基化程度较高。对于其他病毒，先前已经表明糖基化显著影响抗体反应和中和抗体水平。我们推测，SARS-CoV-2的尖峰S的糖基化和糖型对其人类致病特性有严重影响，因此需要考虑以支持未来改进疫苗和新疗法的开发。本项目的目标是在昆虫细胞中生产重组SARS-CoV-2Spike S，并利用MPI开发的合成糖生物技术平台来产生差异糖基化的Spike S和脱糖基化的Spike S。该平台由级联的重组糖基转移酶组成，使我们能够在一锅反应中定制蛋白质的N-糖基化。我们的项目合作伙伴-马格德堡奥托-冯-格里克大学将研究不同糖型的Spike S蛋白与ACE2结合的情况和小鼠的免疫反应。

The ongoing corona virus disease 2019 (Covid-19) pandemic is caused by the coronavirus Sars-CoV-2. Viral entry into human host cells is mediated by the glycoprotein Spike S that binds to angiotensin-converting enzyme 2 (ACE2). Compared to other viral membrane proteins Spike S is heavily N-glycosylated. For other viruses, it has been shown previously that glycosylation significantly impacts antibody response and neutralizing antibody levels. We hypothesize that the glycosylation and the glycoforms of the Spike S of Sars-CoV-2 has a severe impact on its human-pathogenic features and consequently needs to be considered to support future development of improved vaccines and novel therapeutics. Objective of the project is to produce recombinant Sars-CoV-2 Spike S in insect cells and use the synthetic glycobiotechnology platform developed at the MPI to generate differentially glycosylated Spike S as well as deglycosylated Spike S. The platform consists of cascades of recombinant glycosyltransferase that enables us to tailor the N-glycosylation of protein in one-pot reactions. The Spike S proteins with different glycoforms will be investigated for binding to ACE 2 and the immune response in mice by our project partner - the Otto-von-Guericke-University Magdeburg.