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Sortase-Modified Cholera Toxoids Show Specific Golgi Localization

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

in Aminoacyltransferases - genetics / Aminoacyltransferases - metabolism / Animals / Bacterial Proteins - genetics / Bacterial Proteins - metabolism / Biological monitoring / cellular imaging / Cholera / Cholera toxin / Cholera Toxin - metabolism / Chromatography / Cysteine Endopeptidases - metabolism / Endocytosis / Enzymes / Golgi apparatus / Golgi Apparatus - metabolism / Golgi body / Gram-positive bacteria / Humans / Labeling / Lipid rafts / Lipids / Localization / Mammalian cells / Methods / Peptides / Physiological aspects / protein labeling / Proteins / Sortase / Toxins / Toxoids / Tracing / Waterborne diseases

2024

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2024

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2024

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Journal Article

Sortase-Modified Cholera Toxoids Show Specific Golgi Localization

Stevenson, Charlotte M. E.,

Machin, Darren C.,

Andrews, Benjamin I.,

Ungar, Daniel,

Williamson, Daniel J.,

Macdonald, Andrew,

Arnott, Zoe L. P.,

Miller, Victoria J.,

Wasson, Christopher W.,

Webb, Michael E.,

Turnbull, W. Bruce,

Wildsmith, Gemma C.,

Ross, James F.,

Fisher, Peter

2024

Overview

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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Publisher

MDPI AG,MDPI

Subject

Aminoacyltransferases - genetics

/ Aminoacyltransferases - metabolism

/ Animals

/ Bacterial Proteins - genetics

/ Bacterial Proteins - metabolism

/ Biological monitoring

/ cellular imaging