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Sortase-Modified Cholera Toxoids Show Specific Golgi Localization

preprint
submitted on 29.07.2019 and posted on 29.07.2019 by Darren Machin, Daniel Williamson, Peter Fisher, victoria miller, Gemma Wildsmith, James Ross, Christopher Wasson, Andrew MacDonald, Benjamin I. Andrews, Daniel Ungar, W. Bruce Turnbull, Michael Webb
Cholera toxoid is an established tool for use in cellular tracing in neuroscience and cell biology. We use a sortase-labelling approach to generate site-specifically 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 unexpected localization in the medial/trans Golgi. This study suggests a future role for specifically-labelled cholera toxoids in live-cell imaging beyond their current applications in neuronal tracing and labelling of lipid-rafts in fixed cells.

Funding

BB/J014443/1

Biotechnology and Biological Sciences Research Council

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MR/J000604/1

Designing protein-protein interfaces in Cholera Toxin for synthetic virus-like capsid construction

Wellcome Trust

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BB/M005666/1

MR/K012665

MR/S001697/1

BB/P028152/1

BB/R005540/1

History

Email Address of Submitting Author

m.e.webb@leeds.ac.uk

Institution

University of Leeds

Country

United Kingdom

ORCID For Submitting Author

0000-0003-3574-4686

Declaration of Conflict of Interest

The authors do not declare any conflicts of interest

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