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  • Sortase-Modified Cholera To...
    Machin, Darren C; Williamson, Daniel J; Fisher, Peter; Miller, Victoria J; Arnott, Zoe L P; Stevenson, Charlotte M E; Wildsmith, Gemma C; Ross, James F; Wasson, Christopher W; Macdonald, Andrew; Andrews, Benjamin I; Ungar, Daniel; Turnbull, W Bruce; Webb, Michael E

    Toxins, 04/2024, Letnik: 16, Številka: 4
    Journal Article

    Cholera toxoid is an established tool for use in cellular tracing in neuroscience and cell biology. We use a sortase labeling approach to generate site-specific N-terminally modified variants of both the A2-B heterohexamer and B pentamer forms of the toxoid. Both forms of the toxoid are endocytosed by GM1-positive mammalian cells, and while the heterohexameric toxoid was principally localized in the ER, the B pentamer showed an unexpectedly specific localization in the medial/trans-Golgi. This study suggests a future role for specifically labeled cholera toxoids in live-cell imaging beyond their current applications in neuronal tracing and labeling of lipid rafts in fixed cells.