Inhibition of coronavirus release and mitigate COVID-19 pathogenesis

抑制冠状病毒释放并减轻 COVID-19 发病机制

• 批准号: 10272259
• 负责人: PETER D. SUN
• 金额: $ 14.33万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10272259
• 关键词: 2019-nCoV; Affect; Affinity; Antiviral Agents; Binding; Binding Proteins; Biochemistry; Blood Pressure; Bradykinin; CASP1 gene; COVID-19; Cell surface; Clinical; Coronavirus; Coronavirus spike protein; Disease; Drug Targeting; Equilibrium; HIV; HIV Envelope Protein gp120; Human; Infection; Journals; Lectin Receptors; Manuscripts; Mediating; Metalloproteases; Neuraminidase; Oseltamivir; Pathogenesis; Physiological; Polysaccharides; Process; Proteins; Publications; Receptor Inhibition; Recombinants; Severities; Sialic Acids; Site; System; Therapeutic; Viral; glycosylation; influenzavirus; nanomolar; pandemic disease; peptide analog; receptor

To characterize the SARS-CoV-2 spike protein interaction with lectin receptors, we have expressed recombinant SARS-CoV-2 spike prefusion trimer using 293F freestyle expression system. Given the nanomolar high affinity of the spike protein binding to human ACE2, the entry receptor for SARS-CoV-2, and the physiological importance of ACE2 in balancing blood pressure, we investigated if this interaction would affect the enzymatic activity of ACE2. Surprisingly, SARS-CoV-2 trimeric spike protein increased ACE2 proteolytic activity 3-10 fold when fluorogenic caspase-1 substrate and Bradykinin-analog peptides were used to characterize ACE2 activity. In addition, the enhancement was mediated by ACE2 binding of RBD domain of SARS-CoV-2 spike. These results highlighted the altered activity of ACE2 during SARS-CoV-2 infection and would shed new lights on the pathogenesis of COVID-19 and its complications for better treatments. A manuscript describing this finding is under revision for publication in Journal of Biological Chemistry. In addition, we begin to construct a pseudo-coronavirus with SARS-CoV-2 spike envelope. We are in the process to express several lectin receptors either recombinantly or on cell surface.

为了表征 SARS-CoV-2 刺突蛋白与凝集素受体的相互作用，我们使用 293F 自由式表达系统表达重组 SARS-CoV-2 刺突预融合三聚体。 鉴于刺突蛋白与人 ACE2（SARS-CoV-2 的进入受体）结合的纳摩尔高亲和力，以及 ACE2 在平衡血压方面的生理重要性，我们研究了这种相互作用是否会影响 ACE2 的酶活性。令人惊讶的是，当使用荧光 caspase-1 底物和缓激肽类似肽来表征 ACE2 活性时，SARS-CoV-2 三聚体刺突蛋白使 ACE2 蛋白水解活性增加了 3-10 倍。 此外，这种增强是通过 ACE2 结合 SARS-CoV-2 刺突的 RBD 结构域介导的。这些结果强调了 SARS-CoV-2 感染期间 ACE2 活性的改变，并将为了解 COVID-19 的发病机制及其并发症提供新的线索，以提供更好的治疗方法。描述这一发现的手稿正在修订中，以便在《生物化学杂志》上发表。 此外，我们开始构建带有 SARS-CoV-2 刺突包膜的伪冠状病毒。 我们正在重组或在细胞表面表达几种凝集素受体。