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

doi:10.3390/toxins16040194

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

Biology

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Article number	194
Number of pages	13
Journal	Toxins
Volume	16
Issue number	4
DOIs	https://doi.org/10.3390/toxins16040194
Publication status	Published - 16 Apr 2024

Bibliographical note

Keywords

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

Access to Document

10.3390/toxins16040194Licence: CC BY

toxins-16-00194-v3
© 2024 by the authors.
Final published version, 4.68 MBLicence: CC BY

Cite this

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title = "Sortase-Modified Cholera Toxoids Show Specific Golgi Localization",

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.",

keywords = "Humans, Cholera Toxin/metabolism, Cysteine Endopeptidases/metabolism, Golgi Apparatus/metabolism, Animals, Bacterial Proteins/metabolism, Aminoacyltransferases/metabolism, Endocytosis",

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

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doi = "10.3390/toxins16040194",

language = "English",

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T1 - Sortase-Modified Cholera Toxoids Show Specific Golgi Localization

AU - Machin, Darren C

AU - Williamson, Daniel J

AU - Fisher, Peter

AU - Miller, Victoria J

AU - Arnott, Zoe L P

AU - Stevenson, Charlotte M E

AU - Wildsmith, Gemma C

AU - Ross, James F

AU - Wasson, Christopher W

AU - Macdonald, Andrew

AU - Andrews, Benjamin I

AU - Ungar, Daniel

AU - Turnbull, W Bruce

AU - Webb, Michael E

PY - 2024/4/16

Y1 - 2024/4/16

N2 - 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.

AB - 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.

KW - Humans

KW - Cholera Toxin/metabolism

KW - Cysteine Endopeptidases/metabolism

KW - Golgi Apparatus/metabolism

KW - Animals

KW - Bacterial Proteins/metabolism

KW - Aminoacyltransferases/metabolism

KW - Endocytosis

U2 - 10.3390/toxins16040194

DO - 10.3390/toxins16040194

M3 - Article

C2 - 38668619

SN - 2072-6651

VL - 16

JO - Toxins

JF - Toxins

IS - 4

M1 - 194

ER -