Vankadari Naveen, Ketavarapu Vijayasarathy, Mitnala Sasikala, Vishnubotla Ravikanth, Reddy Duvvur Nageshwar, Ghosal Debnath
Monash Biomedicine Discovery Institute, Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria 3800, Australia.
Institute of Translational Research, Department of Genomics and Molecular Biology, Asian Institute of Gastroenterology, Gachibowli, Hyderabad 500032, Telangana, India.
J Phys Chem Lett. 2022 Jun 8:5324-5333. doi: 10.1021/acs.jpclett.2c00967.
The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected more than 520 million people around the globe resulting in more than 6.2 million as of May 2022. Understanding the cell entry mechanism of SARS-CoV-2 and its entire repertoire is a high priority for developing improved therapeutics. The SARS-CoV-2 spike glycoprotein (S-protein) engages with host receptor ACE2 for adhesion and serine proteases furin and TMPRSS2 for proteolytic activation and subsequent entry. Recent studies have highlighted the molecular details of furin and S-protein interaction. However, the structural and molecular interplay between TMPRSS2 and S-protein remains enigmatic. Here, using biochemical, structural, computational, and molecular dynamics approaches, we investigated how TMPRSS2 recognizes and activates the S-protein to facilitate viral entry. First, we identified three potential TMPRSS2 cleavage sites in the S2 domain of S-protein (S2', T1, and T2) and reported the structure of TMPRSS2 with its individual catalytic triad. By employing computational modeling and structural analyses, we modeled the macromolecular structure of TMPRSS2 in complex with S-protein, which incited the mechanism of S-protein processing or cleavage for a new path of viral entry. On the basis of structure-guided drug screening, we also report the potential TMPRSS2 inhibitors and their structural interaction in blocking TMPRSS2 activity, which could impede the interaction with the spike protein. These findings reveal the role of TMPRSS2 in the activation of SARS-CoV-2 for its entry and insight into possible intervention strategies.
由严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的2019冠状病毒病（COVID-19）大流行已感染全球超过5.2亿人，截至2022年5月，导致超过620万人死亡。了解SARS-CoV-2的细胞进入机制及其全部功能是开发改进疗法的当务之急。SARS-CoV-2刺突糖蛋白（S蛋白）与宿主受体血管紧张素转换酶2（ACE2）结合以实现粘附，并与丝氨酸蛋白酶弗林蛋白酶和跨膜丝氨酸蛋白酶2（TMPRSS2）结合以进行蛋白水解激活并随后进入细胞。最近的研究突出了弗林蛋白酶和S蛋白相互作用的分子细节。然而，TMPRSS2与S蛋白之间的结构和分子相互作用仍然是个谜。在这里，我们使用生物化学、结构、计算和分子动力学方法，研究了TMPRSS2如何识别和激活S蛋白以促进病毒进入。首先，我们在S蛋白的S2结构域中确定了三个潜在的TMPRSS2切割位点（S2'、T1和T2），并报告了TMPRSS2及其单个催化三联体的结构。通过采用计算建模和结构分析，我们对TMPRSS2与S蛋白复合物的大分子结构进行了建模，这引发了S蛋白加工或切割的机制，从而为病毒进入开辟了一条新途径。基于结构导向的药物筛选，我们还报告了潜在的TMPRSS2抑制剂及其在阻断TMPRSS2活性方面的结构相互作用，这可能会阻碍与刺突蛋白的相互作用。这些发现揭示了TMPRSS2在激活SARS-CoV-2进入细胞中的作用，并为可能的干预策略提供了见解。
