A split luciferase system for studying coronavirus M^pro dimerization in vitro and in living cells.

ABSTRACT: The main protease enzyme (M^pro) of coronaviruses cleaves the viral polyprotein into functional units essential for virus replication. Prior work has demonstrated that M^pro functions as a homodimer. However, studies on the mechanism of dimerization have been challenging because the purified protease is mostly dimeric, dimerization-defective mutants lack proteolytic activity, and robust cell-based assays have yet to be reported. To enable work on M^pro dimerization, we have developed a quantitative luciferase-based SARS-CoV-2 (SARS2) M^pro biosensor that accurately reports protein dimerization in living cells and, upon purification, also in vitro. Co-transfection of cells with a construct expressing M^pro fused to the 18 kDa LargeBiT of luc

SUBMITTER: Delgado R 

PROVIDER: S-EPMC12719657 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature