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Proteins, Expression, Isolation and Analysis> | The cell culture supernatants or cell lysates were prepared as reported previously [7]. For immunoblotting, the samples were mixed with SDS-PAGE buffer and heated at 100 °C for 10 min, followed by electrophoresis in 12% polyacrylamide gel (GenScript) | Get A Quote |
Bacillus cereus is a clinically significant foodborne pathogen that causes severe gastrointestinal and non-gastrointestinal disease. Cereolysin O (CLO) is a putative virulence factor of B. cereus, and its function remains to be investigated. In this study, we examined the biological activity of CLO from a deep sea B. cereus isolate. CLO was highly toxic to mammalian cells and triggered pyroptosis through NLRP3 inflammasome-mediated caspase 1 and gasdermin D activation. CLO-induced cell death involved ROS accumulation and K+ efflux, and was blocked by serum lipids. CLO bound specifically to cholesterol, and this binding was essential to CLO cytotoxicity. The structural integrity of the three tryptophan residues ... More