A covalent chemical probe for Chikungunya nsP2 cysteine protease with antialphaviral activity and proteome-wide selectivity
Abstract
Abstract Chikungunya is a mosquito-borne viral disease that causes fever and severe joint pain for which there is no direct acting drug treatments. Vinyl sulfone SGC-NSP2PRO-1 (3) was identified as a potent inhibitor of the nsP2 cysteine protease (nsP2pro) that reduced viral titer against infectious isolates of Chikungunya and other alphaviruses. The covalent warhead in 3 captured the active site C478 and inactivated nsP2pro with a k inact/K i ratio of 5950 M–1 s–1. The vinyl sulfone 3 was inactive across a panel of 23 other cysteine proteases and demonstrated remarkable proteome-wide selectivity by two chemoproteomic methods. A negative control analog SGC-NSP2PRO-1N (4) retained the isoxazole core and covalent warhead but demonstrated > 100-fold decrease in enzyme inhibition. Both 3 and 4 were stable across a wide range of pH in solution and upon prolonged storage as solids. Vinyl sulfone 3 and its negative control 4 will find utility as high-quality chemical probes to study the role of the nsP2pro in cellular studies of alphaviral replication and virulence.
Article Details
Authors (27)
Anirban Ghoshal
Edwin G. Tse
Mohammad Anwar Hossain
Kesatebrhan Haile Asressu
Eric M. Merten
John D. Sears
Department of Microbiology and Immunology, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill
Stefanie Howell
Sumera Perveen
Jane Burdick
Noah L. Morales
Sabian A. Martinez
Isabella Law
Bennett J. Davenport
Thomas E. Morrison
Zachary J. Streblow
Daniel N. Streblow
Angie L. Mordant
Thomas S. Webb
Aurora Cabrera
Laura E. Herring
Department of Pharmacology, University of North Carolina Metabolomics and Proteomics Core, University of North Carolina at Chapel Hill
Cheryl H. Arrowsmith
Kenneth H. Pearce
Nathaniel J. Moorman
Department of Microbiology and Immunology, University of North Carolina at Chapel Hill
Mark T. Heise
Rafael M. Couñago
Peter J. Brown
Timothy M. Willson