Biophysical and immunoinformatics insights into TolC, an outer membrane multidrug efflux channel of Vibrio cholerae

Vibrio cholerae is a gram-negative bacterium known for causing severe diarrhea and disrupting the barrier function of intestinal cells. Its outer membrane proteins (OMPs), particularly TolC, are crucial for maintaining bacterial homeostasis and pathogenicity. TolC is vital for antibiotic resistance and molecular efflux, making it a key target for study. However, extracting large quantities of soluble OMPs is challenging. To address this, TolC from V. cholerae is refolded in vitro using detergents and lipids and characterized biophysically. Additionally, its antigenic potential is explored through immunoinformatics. The results show that different folding reagents vary in effectiveness, with some promoting a near-native quaternary structure, specifically a trimer held together by non-covalent interactions. Moreover, TolC has surface-exposed epitopes capable of triggering an immune response. Owing to its role in antimicrobial resistance, this study provides valuable structural insights into refolded TolC in various detergents and lipids, potentially aiding large-scale production of structurally accurate protein for drug and vaccine development.