Abstract
RECOGNITION of the oligosaccharide portion of ganglioside GM1 in membranes of target cells by the heat-labile enterotoxin from Escherichia coli is the crucial first step in its pathogenesis, as it is for the closely related cholera toxin1–3. These toxins have five B subunits, which are essential for GM1 binding, and a single A subunit, which needs to be nicked by proteolysis and reduced, yielding an Al–'enzyme' and an A2–'linker' peptide. A1 is translocated across the membrane of intestinal epithelial cells, possibly after endocytosis4,5, upon which it ADP-ribosylates the G protein Gsa (reviewed in refs 2, 3, 6). The mechanism of binding and translocation of these toxins has been extensively investigated1,2,7–20, but how the protein is orientated on binding is still not clear10–12,18. Knowing the precise arrangement of the ganglioside binding sites of the toxins will be useful for designing drugs against the diarrhoeal diseases caused by organisms secreting these toxins and in the development of oral vaccines against them21,22. We present here the three-dimensional structure of the E. coli heat-labile enterotoxin complexed with lactose. This reveals the location of the binding site of the terminal galactose of GM1, which is consistent with toxin binding to the target cell with its A1 fragment pointing away from the membrane. A small helix is identified at the carboxy terminus of A2 which emerges through the central pore of the B subunits and probably comes into contact with the membrane upon binding, whereas the A1 subunit is flexible with respect to the B pentamer.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Eidels, L., Proia, R. L. & Hart, D. A. Microbiol. Rev. 47, 596–620 (1983).
van Heyningen, S. in Current Topics in Membranes and Transport Vol. 18, 445–471 (Academic, New York, 1983).
Moss, J. & Vaughan, M. Adv. Enzym. 61, 303–379 (1988).
Montesano, R., Roth, J., Robert, A. & Orci, L. Nature 296, 651–653 (1982).
Janicot, M., Fouque, F. & Desbuquois, B. J. biol. Chem. 266, 12858–12865 (1991).
Finkelstein, R. A. in Immunochemical and Molecular Genetic Analysis of Bacterial Pathogens (eds Owen, P. & Foster, T. J.) 85–102 (Elsevier, New York, 1988).
Sattler, J., Schwarzmann, G., Staerk, J., Ziegler, W. & Wiegandt, H. Hoppe Seyler's Z. physiol. Chem. 358, 159–163 (1977).
Clements, J. D. & Finkelstein, R. A. Infect. Immunity 24, 760–769 (1979).
De Wolf, M. J. S., Fridkin, M. & Kohn, L. D. J. biol. Chem. 256, 5489–5496 (1981).
Tomasi, M. & Montecucco, C. J. biol. Chem. 256, 11177–11181 (1981).
Wisnieski, B. J. & Bramhall, J. S. Nature 289, 319–321 (1981).
Dwyer, J. D. & Bloomfield, V. A. Biochemistry 21, 3227–3231 (1982).
Holmgren, J., Fredman, P., Lindblad, M., Svennerholm, A.-M. & Svennerholm, L. Infect. Immunity 38, 424–433 (1982).
Ludwig, D. S., Holmes, R. K. & Schoolnik, G. K. J. biol. Chem. 260, 12528–12534 (1985).
Tsuji, T., Honda, T., Miwatani, T., Wakabayashi, S. & Matsubara, H. J. biol. Chem. 260, 8552–8558 (1985).
Griffiths, S. L., Finkelstein, R. A. & Critchley, D. R. Biochem. J. 238, 313–322 (1986).
Fukuta, S., Magnani, J. L., Twiddy, E. M., Holmes, R. K. & Ginsburg, V. Infect. Immunity 56, 1748–1753 (1988).
Ribi, H. A., Ludwig, D. S., Mercer, K. L., Schoolnik, G. K. & Kornberg, R. D. Science 239, 1272–1276 (1988).
Goins, B. & Freire, E. Biochemistry 27, 2046–2052 (1988).
Surewicz, W. K., Leddy, J. J. & Mantsch, H. H. Biochemistry 29, 8106–8111 (1990).
Svennerholm, A.-M., Holmgren, J., Sack, D. A. & Bardhan, P. K. Lancet i, 305–307 (1982).
de Aizpurua, H. J. & Russell-Jones, G. J. J. exp. Med. 167, 440–451 (1988).
Vyas, N. K., Vyas, M. N. & Quiocho, F. A. J. biol. Chem. 266, 5226–5237 (1991).
Rutenber, E. & Robertus, J. D. Proteins 10, 260–269 (1991).
Sixma, T. K. et al. Nature 351, 371–377 (1991).
McDaniel, R. V. & McIntosh, T. J. Biophys. J. 49, 94–96 (1986).
Pelham, H. R. B. EMBO J. 8, 3171–3176 (1989).
Arnone, A. et al. J. biol. Chem. 246, 2302–2316 (1971).
Read, R. J. Acta crystallogr. A42, 140–149 (1986).
Leong, J., Vinal, A. C. & Dallas, W. S. Infect. Immunity 48, 73–77 (1985).
Pronk, S. E. et al. J. biol. Chem. 260, 13580–13584 (1985).
Tronrud, D. E., Ten Eyck, L. F. & Matthews, B. W. Acta crystallogr. A43, 489–501 (1987).
Brünger, A. T., Kuriyan, J. & Karplus, M. Science 235, 458–460 (1987).
Lesk, A. M. & Hardman, K. D. Meth. Enzym. 115, 381–390 (1985).
Priestle, J. P. J. appl. Crystallogr. 21, 572–576 (1988).
Sanchez, J., Hirst, T. R. & Uhlin, B. E. Gene 64, 265–275 (1988).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sixma, T., Pronk, S., Kalk, K. et al. Lactose binding to heat-labile enterotoxin revealed by X-ray crystallography. Nature 355, 561–564 (1992). https://doi.org/10.1038/355561a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/355561a0
This article is cited by
-
Holotoxin disassembly by protein disulfide isomerase is less efficient for Escherichia coli heat-labile enterotoxin than cholera toxin
Scientific Reports (2022)
-
Enhancement of toxin- and virus-neutralizing capacity of single-domain antibody fragments by N-glycosylation
Applied Microbiology and Biotechnology (2009)
-
Structure of benzyl T-antigen disaccharide bound to Amaranthus caudatus agglutinin
Nature Structural Biology (1997)
-
Interaction of human immunodeficiency virus type 1 envelope protein with liposomes containing galactosylceramide
Perspectives in Drug Discovery and Design (1996)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.