一种具有抗α病毒活性和全蛋白质组选择性的nsP2半胱氨酸蛋白酶的共价化学探针。
A Covalent Chemical Probe for nsP2 Cysteine Protease with Antialphaviral Activity and Proteome-wide Selectivity.
作者信息
Ghoshal Anirban, Tse Edwin G, Hossain Mohammad Anwar, Asressu Kesatebrhan Haile, Merten Eric M, Sears John D, Howell Stefanie, Perveen Sumera, Burdick Jane, Morales Noah L, Martinez Sabian A, Law Isabella, Davenport Bennett J, Morrison Thomas E, Streblow Zachary J, Streblow Daniel N, Mordant Angie L, Webb Thomas S, Cabrera Aurora, Herring Laura E, Arrowsmith Cheryl H, Pearce Kenneth H, Moorman Nathaniel J, Heise Mark T, Couñago Rafael Miguez, Brown Peter J, Willson Timothy M
机构信息
Structural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
READDI AViDD Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
出版信息
Res Sq. 2024 Nov 12:rs.3.rs-5363451. doi: 10.21203/rs.3.rs-5363451/v1.
RA-0003022 () was identified as a high-quality covalent chemical probe for nsP2 cysteine protease (nsP2pro). Isoxazole covalently captured the active site C478 and inactivated the enzyme with a / ratio of 6000 Ms. A negative control analog RA-0025453 () retained the covalent warhead but demonstrated >100-fold decrease in enzyme inhibition. Isoxazoles and were stable across a wide range of pH in solution and upon prolonged storage as solids. The covalent chemical probe was inactive across a panel of 23 human and viral cysteine proteases and demonstrated remarkable proteome-wide selectivity by two chemoproteomic methods. Isoxazole reduced viral titer against infectious isolates of , , and Venezuelan Equine Encephalitis viruses demonstrating its activity on both New and Old World alphaviruses. Isoxazole and its negative control will find utility as covalent chemical probes to study the role of the nsP2pro in alphaviral replication and virulence.
RA - 0003022（）被鉴定为一种用于nsP2半胱氨酸蛋白酶（nsP2pro）的高质量共价化学探针。异恶唑共价捕获活性位点C478并使该酶失活，解离常数/抑制常数（/）为6000 M⁻¹s⁻¹。阴性对照类似物RA - 0025453（）保留了共价弹头，但酶抑制作用降低了100倍以上。异恶唑和在溶液中广泛的pH范围内以及作为固体长期储存时都很稳定。共价化学探针在一组23种人类和病毒半胱氨酸蛋白酶中无活性，并通过两种化学蛋白质组学方法显示出显著的全蛋白质组选择性。异恶唑降低了针对辛德毕斯病毒、基孔肯雅病毒和委内瑞拉马脑炎病毒感染性分离株的病毒滴度，证明了其对新旧世界甲病毒均有活性。异恶唑及其阴性对照将作为共价化学探针用于研究nsP2pro在甲病毒复制和毒力中的作用。
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