新型冠状病毒主蛋白酶M49L及T93I突变体多维生化性质表征  

Multiscale Characterization of M49L and T93I Mutations to the Biochemical Properties of the Main Protease of SARS-CoV-2

作　　者：杨润泽 王莉 陈聪聪 潘朋朋 孙佳聪 陈成 YANG Runze;WANG Li;CHEN Congcong;PAN Pengpeng;SUN Jiacong;CHEN Cheng(School of Life Science,Tianjin University,Tianjin 300000,China;Institute of Clinical Pharmacology,Anhui Medical University,Hefei 230032,China)

机构地区：[1]天津大学生命科学学院,天津300000 [2]安徽医科大学临床药理研究所,合肥230032

出　　处：《病毒学报》2024年第6期1230-1237,共8页Chinese Journal of Virology

摘　　要：2019年出现的新型冠状病毒(SARS-CoV-2)对人类的健康造成了持续性的威胁。在新冠病毒编码的众多蛋白中,主蛋白酶(Main protease,M^(pro))由于其酶切功能的重要性、高度的保守性以及在人体内无同源蛋白的优势,成为了当前抗新冠药物开发的核心靶点之一。然而,随着病毒持续进化,M^(pro)出现了众多突变,这些突变在各结构域上分布广泛,影响M^(pro)各种性质,从而促进了SARS-CoV-2的持续进化。本研究选取了活性中心的M49L以及远离活性中心的T93I两个天然高频突变进行包括蛋白聚集状态、酶动力学以及热稳定性等生化性质的表征,发现M49L和T93I突变对聚集状态和热稳定性无明显影响,对M^(pro)催化效率分别提高了99%以及12%。本研究成功揭示两个突变对M^(pro)生化性质的影响,为后续抗新冠药物开发提供了指导。The emergence of severe acute respiratory syndrome-coronavirus-2(SARS-CoV-2)in 2019 has posed a persistent threat to human health.Among the various proteins encoded by SARS-CoV-2,the main protease(M^(pro))is one of the most crucial targets for drug development,due to its dominant role in maturation of non-structural proteins,high conservation,and absence of homologous proteins in humans.However,numerous mutations have occurred in M^(pro).These mutations distributes across various structural domains and affect the properties of M^(pro),thereby contributing to the ongoing evolution of SARS-CoV-2.M49L and T93I,two naturally occurring high-frequency mutations which locate within and outside of the active sites of M^(pro),respectively,were subjected to assessment the state of protein oligomerization,enzyme kinetic activity,and thermal stability.Catalytic efficiency(rather than the protein oligomerization state or thermal stability)was increased significantly for M49L and T93I mutants,with increases of 99%and 12%,respectively.We clarified the biochemical properties influenced by these mutations and,thus,offer valuable insights for further development of anti-SARS-CoV-2 drug.

关 键 词：新型冠状病毒 主蛋白酶 天然突变 聚集状态 酶动力学 热稳定性 

分 类 号：R373.1[医药卫生—病原生物学]