Imaging the intracellular trafficking and state of the AB5 quaternary structure of cholera toxin

Philippe I.H. Bastiaens, Irina V. Majoul*, Peter J. Verveer, Hans Dieter Söling, Thomas M. Jovin

*Corresponding author for this work
232 Citations (Scopus)

Abstract

The subcellular localization and corresponding quaternary state of fluorescent labelled cholera toxin were determined at different time points after exposure to living cells by a novel form of fluorescence confocal microscopy. The compartmentalization and locus of separation of the pentameric B subunits (CTB) from the A subunit (CTA) of the toxin were evaluated on a pixel-by-pixel (voxel-by-voxel) basis by measuring the fluorescence resonance energy transfer (FRET) between CTB labelled with the sulfoindocyanine dye Cy3 and an antibody against CTA labelled with Cy5. The FRET efficiency was determined by a new technique based on the release of quenching of the Cy3 donor after photodestruction of the Cy5 acceptor in a region of interest within the cell. The results demonstrate vesicular transport of the holotoxin from the plasma membrane to the Golgi compartment with subsequent separation of the CTA and CTB subunits. The CTA subunit is redirected to the plasma membrane by retrograde transport via the endoplasmic reticulum whereas the CTB subunit persists in the Golgi compartment.

Original languageEnglish
JournalEMBO Journal
Volume15
Issue number16
Pages (from-to)4246-4253
Number of pages8
ISSN0261-4189
DOIs
Publication statusPublished - 15.08.1996

Research Areas and Centers

  • Academic Focus: Center for Infection and Inflammation Research (ZIEL)

Fingerprint

Dive into the research topics of 'Imaging the intracellular trafficking and state of the AB5 quaternary structure of cholera toxin'. Together they form a unique fingerprint.

Cite this