TurboID-mediated surface protein biotinylation to inhibit the growth ofStaphylococcus aureus

Abstract

AbstractStaphylococcus aureus(S. aureus) infection is major cause of nosocomial infections. Antibiotic treatment forS. aureusremains the primary solution for managingS. aureusinfections, which, however, increases the risk of antibiotic resistance. To broaden the resolutions onS. aureusinfection, here we report TurboID-mediated protein proximity technologies to inhibit the growth ofS. aureus. To achieve this goal, we utilized synthetic biology techniques to create a fusion protein named N-AgrD-TurboID (Agr-ID). The N-AgrD domain includes auto-inducer peptide (AIP) which combined to the surface AgrC protein onS. aureus. As such, TurboID then catalyzed the production of biotinoyl-5’-AMP anhydride, triggering the biotinylation of surface proteins onS. aureus25923 which were visualized by using fluorescence microscopy after incubating with Alexa Fluor 647-conjugated streptavidin. The biotinylation of surface protein onS. aureus25923,S. aureus43300, andS. aureus6538 (MRSA) also resulted in growth inhibition and impaired colonization. Moreover, the biotinylation on surface protein further inhibited virulence protein production inS. aureus25923, as indicated by reduced apoptosis of HEK 293T cells after treatment withS. aureus25923 lysates. Overall, our work reveals that the biotinylation of surface proteins can inhibit the growth and toxicity ofS. aureus25923,S. aureus43300, andS. aureus6538 (MRSA), indicating therapeutic potential in clinical treatment.