Sortase-Modified Cholera Toxoids Show Specific Golgi Localization

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

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish
Article number194
Number of pages13
Issue number4
Publication statusPublished - 16 Apr 2024

Bibliographical note

© 2024 by the authors.


  • Humans
  • Cholera Toxin/metabolism
  • Cysteine Endopeptidases/metabolism
  • Golgi Apparatus/metabolism
  • Animals
  • Bacterial Proteins/metabolism
  • Aminoacyltransferases/metabolism
  • Endocytosis

Cite this