Optimization of quenched fluorescent peptide substrates of SARS-CoV-2 3CL^pro main protease (Mpro) from proteomic identification of P6-P6' active site specificity.

ABSTRACT: SARS-CoV-2 3C-like main protease (3CL^pro) is essential for protein excision from the viral polyprotein. 3CL^pro inhibitor drug development to block SARS-CoV-2 replication focuses on the catalytic non-prime (P) side for specificity and potency, but the importance of the prime (P') side in substrate specificity and for drug development remains underappreciated. We determined the P6-P6' specificity for 3CL^pro from >800 cleavage sites that we identified using Proteomic Identification of Cleavage site Specificity (PICS). Cleavage occurred after the canonical P1-Gln and non-canonical P1-His and P1-Met residues. Moreover, P3 showed a preference for Arg/Lys and P3' for His. Essential H-bonds between the N-terminal Ser1 of protomer-B in 3CL^pro dimers form

SUBMITTER: Cesar Ramos de Jesus H 

PROVIDER: S-EPMC11237654 | biostudies-literature | 2024 Jun

REPOSITORIES: biostudies-literature