AB5 bacterial toxins relevant to human disease, Cholera toxin and Shiga

toxin, as model cargo for intracellular membrane trafficking studies

    Cholera Toxin (CTX), an AB5 exotoxin produced by pathogenic Vibrio cholerae bacteria, causes disease with pandemic potential in humans. Symptoms include diarrheria, severe dehydration, and if kidneys are affected blood in the urine. Left untreated this disease has a high mortality rate, but is treatable when recognized in time.


    CTX binds to its receptor, glycosphingolipid GM1, at the plasma membrane surface of gut and kidney cells. Following internalization in endosomes CTX arrives at the Golgi complex. We have shown that Golgi-to-ER transport of CTX is crucially dependent on COP I proteins (Majoul et al. 1996, 1998, 2001).


    In the lab we routinely use fluorescently labeled CTX-K63 (a non-toxic CTX mutant) as well as Shiga toxin (purified from Shigella dysenteriae) as model cargos for in vivo trafficking (Majoul et al., 2002, 2006; Frigerio et al., 2007).

CTX fills up the Golgi complex ~ 30 min after internalization in Vero kidney cells. The Golgi complex and CTX -  loaded transport vesicles are seen, which travel to and from the Golgi along microtubule tracks. This time lapse movie plays at 25x real speed, total recording time ~17 min (1000 frames). A 63x NA 1.4 oil immersion objective was used.  Size bar: 10 micron.

CHOLERA TOXIN INTERNALIZATION IN LIVING CELLS

     SELECTED LAB PUBLICATIONS RELATED TO THIS TOPIC:


Majoul I, Gao L, Betzig E, Onichtchouk D, Butkevich E, Kozlov Y, Bukauskas F, Bennett MVL, Lippincott-Schwartz J, Duden R (2013). Fast structural responses of gap junction membrane domains to AB5 toxins. PROC. NATL. ACAD. SCI. USA 110(44): E4125-4133.  doi: 10.1073/pnas.1315850110    (link to article)


Berger Z, Davies JE, Pasco M, Majoul I, O’Kane C, Rubinsztein D (2006). Deleterious and protective properties of an aggregate-prone protein with a polyalanine expansion.  HUM. MOL. GENET. 15: 453-465   (link to article)


Majoul I, Jia Y, Duden R (2006). Practical Fluorescence Resonance Energy Transfer   (FRET) or molecular nanobioscopy of living cells. Book chapter in: “Handbook of Biological Confocal Microscopy” 3rd ed. (Pawley, J. (Ed.), pp. 788-808. Springer Internationa(PDF)

    

Majoul I (2004). Analysing the action of bacterial toxins in living cells with fluorescence resonance energy transfer (FRET). Int. J. Med. Microbiol. 293: 495-503  (link to article)


Majoul I, Goroshkov A, Butkevich E, Verrier S, Duden R (2003). Live cell analyses of transport complex formation with FRET, in: “Genetically Engineered and Optical Probes for Biomedical Applications” (Eds. A. Savitsky et al.). Proceedings of the Society for Optical Engineering (SPIE) 4967: 11-18    (link to article)


Bacia K, Majoul I, Schwille P (2002). Probing the endocytic pathway in live cells using dual-color fluorescence

cross-correlation analysis. BIOPHYS. J. 83: 1184-1193  (link to article)


Majoul I, Schmidt T, Pomasanova M, Boutkevich E, Kozlov Y, Söling HD (2002). Differential expression of receptors for Shiga and Cholera toxin is regulated by the cell  cycle.  J. Cell Sci. 115: 817-826  (link to article)


Majoul I, Straub M, Duden R, Hell SW, Söling HD (2002). Fluorescence resonance energy transfer analysis of protein-protein interactions in single living cells by multifocal multiphoton microscopy.  J. Biotechnol. 82: 267-277   (link to article)


Majoul I, Straub M, Hell S, Duden R, Söling HD (2001). KDEL-cargo regulates interactions between proteins involved in COPI vesicle traffic. Measurements in living cells using FRET. DEV. CELL  1: 139-153    COVER: July 2001 issue   [Recommended in F1000Prime] (link to article)


Majoul I, Sohn K, Wieland FT, Pepperkok R, Pizza M, Hillemann J, Söling HD (1998). KDEL receptor (Erd2p)-mediated retrograde transport of the cholera toxin A subunit from the Golgi involves COPI, p23, and the COOH terminus of Erd2p.  J. CELL BIOL. 143: 601-612  (link to article)


Majoul I, Ferrari D, Söling HD (1997). Reduction of protein disulfide bonds in an oxidizing environment. The disulfide bridge of cholera toxin A-subunit is reduced in the endoplasmic reticulum. FEBS Lett. 401: 104-108  

(link to article)


Bastiaens PI, Majoul I, Verveer PJ, Söling HD, Jovin TM (1996). Imaging the intracellular trafficking and state of the AB5 quaternary structure of cholera toxin.  EMBO J. 15: 4246-4253    (link to article)


Majoul I, Bastiaens PI, Söling HD (1996). Transport of an external Lys-Asp-Glu-Leu (KDEL) protein from the plasma membrane to the endoplasmic reticulum: studies with cholera toxin in Vero cells. J. CELL BIOL. 133: 777-789 

(link to article)