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Sortase-Modified Cholera Toxoids Show Specific Golgi Localization.
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.
Affiliation
  • Machin DC; School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Williamson DJ; School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Fisher P; Department of Biology, University of York, York YO10 5DD, UK.
  • Miller VJ; Department of Biochemistry, University of Bristol, Bristol BS8 1QU, UK.
  • Arnott ZLP; School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Stevenson CME; School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Wildsmith GC; School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Ross JF; School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Wasson CW; Faculty of Biological Sciences, Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Macdonald A; Faculty of Biological Sciences, Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Andrews BI; GlaxoSmithKline, Medicines Research Centre, Gunnels Wood Road, Stevenage SG1 2NY, UK.
  • Ungar D; Department of Biology, University of York, York YO10 5DD, UK.
  • Turnbull WB; School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
  • Webb ME; School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
Toxins (Basel) ; 16(4)2024 Apr 16.
Article in En | MEDLINE | ID: mdl-38668619
ABSTRACT
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-B5 heterohexamer and B5 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 B5 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.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Cysteine Endopeptidases / Cholera Toxin / Golgi Apparatus Limits: Animals / Humans Language: En Journal: Toxins (Basel) Year: 2024 Document type: Article
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Cysteine Endopeptidases / Cholera Toxin / Golgi Apparatus Limits: Animals / Humans Language: En Journal: Toxins (Basel) Year: 2024 Document type: Article