Synthetic bulky NS4A peptide variants bind to and inhibit HCV NS3 protease

Moustafa El-Sayed El-Araby, Abdlesattar M Omar, Sameh H Soror, Stefan T Arold, Maan T Khayat, Hani Z Asfour, Faida H Bamane, Mahmoud A El-Faky
Journal of Adv. Res, (2020)

Synthetic bulky NS4A peptide variants bind to and inhibit HCV NS3 protease

Keywords

DSLSFluorescence AnisotropyAllosteric InhibitorHCVNS3/4APeptide Mutants

Abstract

​NS4A is a non-structural multi-tasking small peptide that is essential for HCV maturation and replication. The central odd-numbered hydrophobic residues of NS4A (Val-23’ to Leu-31‘)i are essential for activating NS3 upon NS3/4A protease complex formation. This study aims to design new specific allosteric NS3/4A protease inhibitors by mutating Val-23‘, Ile-25‘, and Ile-29‘ into bulkier amino acids. Pep-15, a synthetic peptide, showed higher binding affinity towards HCV-NS3 subtype-4 than native NS4A. The Kd of Pep-15 (80.0 ± 8.0 nM) was twice as high as that of native NS4A (169 ± 37 nM). The mutant Pep-15 inhibited the catalytic activity of HCV-NS3 by forming an inactive complex. Molecular dynamics simulations suggested that a cascade of conformational changes occurred, especially in the catalytic triad arrangements, thereby inactivating NS3. A large shift in the position of Ser-139 was observed, leading to loss of critical hydrogen bonding with His-57. Even though this study is not a classic drug discovery study—nor do we propose Pep-15 as a drug candidate—it serves as a stepping stone towards developing a potent inhibitor of hitherto untargeted HCV subtypes.

Code

Doi:10.1016/j.jare.2020.01.003

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