Skip to main content
Protein Science : A Publication of the Protein Society logoLink to Protein Science : A Publication of the Protein Society
. 1996 Jun;5(6):1184–1188. doi: 10.1002/pro.5560050621

Tumor marker disaccharide D-Gal-beta 1, 3-GalNAc complexed to heat-labile enterotoxin from Escherichia coli.

F van den Akker 1, E Steensma 1, W G Hol 1
PMCID: PMC2143437  PMID: 8762150

Abstract

Heat-labile enterotoxin (LT) is part of the cholera toxin (CT) family and consists of a catalytic A subunit and a B pentamer that serves to recognize the oligosaccharide part of the GM1 ganglioside receptor. We report here the crystal structure of heat-labile enterotoxin in complex with the disaccharide portion of the Thomsen-Friedenreich (T-antigen) tumor marker. The toxin:carbohydrate complex is determined to 2.13 A resolution, yielding an R-factor of 18.5%. The T-antigen disaccharide, D-Gal-beta 1,3-GalNAc-Ser/Thr, is present in more than 85% of human carcinomas and monitoring its autoimmune response is used for the early detection of tumors. Insight into the molecular recognition of this tumor antigen by sugar binding proteins can benefit the development of a diagnostic tool for human carcinomas as well as a T-antigen directed anticancer drug delivery system.

Full Text

The Full Text of this article is available as a PDF (3.7 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bray J., MacLean G. D., Dusel F. J., McPherson T. A. Decreased levels of circulating lytic anti-T in the serum of patients with metastatic gastrointestinal cancer: a correlation with disease burden. Clin Exp Immunol. 1982 Jan;47(1):176–182. [PMC free article] [PubMed] [Google Scholar]
  2. Brünger A. T., Kuriyan J., Karplus M. Crystallographic R factor refinement by molecular dynamics. Science. 1987 Jan 23;235(4787):458–460. doi: 10.1126/science.235.4787.458. [DOI] [PubMed] [Google Scholar]
  3. Jobling M. G., Holmes R. K. Analysis of structure and function of the B subunit of cholera toxin by the use of site-directed mutagenesis. Mol Microbiol. 1991 Jul;5(7):1755–1767. doi: 10.1111/j.1365-2958.1991.tb01925.x. [DOI] [PubMed] [Google Scholar]
  4. Jones T. A., Zou J. Y., Cowan S. W., Kjeldgaard M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr A. 1991 Mar 1;47(Pt 2):110–119. doi: 10.1107/s0108767390010224. [DOI] [PubMed] [Google Scholar]
  5. Lotan R., Skutelsky E., Danon D., Sharon N. The purification, composition, and specificity of the anti-T lectin from peanut (Arachis hypogaea). J Biol Chem. 1975 Nov 10;250(21):8518–8523. [PubMed] [Google Scholar]
  6. MacLean G. D., Longenecker B. M. Clinical significance of the Thomsen-Friedenreich antigen. Semin Cancer Biol. 1991 Dec;2(6):433–439. [PubMed] [Google Scholar]
  7. Merritt E. A., Murphy M. E. Raster3D Version 2.0. A program for photorealistic molecular graphics. Acta Crystallogr D Biol Crystallogr. 1994 Nov 1;50(Pt 6):869–873. doi: 10.1107/S0907444994006396. [DOI] [PubMed] [Google Scholar]
  8. Merritt E. A., Sarfaty S., Chang T. T., Palmer L. M., Jobling M. G., Holmes R. K., Hol W. G. Surprising leads for a cholera toxin receptor-binding antagonist: crystallographic studies of CTB mutants. Structure. 1995 Jun 15;3(6):561–570. doi: 10.1016/s0969-2126(01)00190-3. [DOI] [PubMed] [Google Scholar]
  9. Merritt E. A., Sixma T. K., Kalk K. H., van Zanten B. A., Hol W. G. Galactose-binding site in Escherichia coli heat-labile enterotoxin (LT) and cholera toxin (CT). Mol Microbiol. 1994 Aug;13(4):745–753. doi: 10.1111/j.1365-2958.1994.tb00467.x. [DOI] [PubMed] [Google Scholar]
  10. Pai L. H., Batra J. K., FitzGerald D. J., Willingham M. C., Pastan I. Anti-tumor activities of immunotoxins made of monoclonal antibody B3 and various forms of Pseudomonas exotoxin. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3358–3362. doi: 10.1073/pnas.88.8.3358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pai L. H., FitzGerald D. J., Tepper M., Schacter B., Spitalny G., Pastan I. Inhibition of antibody response to Pseudomonas exotoxin and an immunotoxin containing Pseudomonas exotoxin by 15-deoxyspergualin in mice. Cancer Res. 1990 Dec 15;50(24):7750–7753. [PubMed] [Google Scholar]
  12. Pastan I., Chaudhary V., FitzGerald D. J. Recombinant toxins as novel therapeutic agents. Annu Rev Biochem. 1992;61:331–354. doi: 10.1146/annurev.bi.61.070192.001555. [DOI] [PubMed] [Google Scholar]
  13. Pereira M. E., Kabat E. A., Lotan R., Sharon N. Immunochemical studies on the specificity of the peanut (Arachis hypogaea) agglutinin. Carbohydr Res. 1976 Oct;51(1):107–118. doi: 10.1016/s0008-6215(00)84040-9. [DOI] [PubMed] [Google Scholar]
  14. Pronk S. E., Hofstra H., Groendijk H., Kingma J., Swarte M. B., Dorner F., Drenth J., Hol W. G., Witholt B. Heat-labile enterotoxin of Escherichia coli. Characterization of different crystal forms. J Biol Chem. 1985 Nov 5;260(25):13580–13584. [PubMed] [Google Scholar]
  15. Schlepper-Schäfer J., Springer G. F. Carcinoma autoantigens T and Tn and their cleavage products interact with Gal/GalNAc-specific receptors on rat Kupffer cells and hepatocytes. Biochim Biophys Acta. 1989 Oct 9;1013(3):266–272. doi: 10.1016/0167-4889(89)90145-6. [DOI] [PubMed] [Google Scholar]
  16. Sixma T. K., Pronk S. E., Kalk K. H., van Zanten B. A., Berghuis A. M., Hol W. G. Lactose binding to heat-labile enterotoxin revealed by X-ray crystallography. Nature. 1992 Feb 6;355(6360):561–564. doi: 10.1038/355561a0. [DOI] [PubMed] [Google Scholar]
  17. Springer G. F., Cheingsong-Popov R., Schirrmacher V., Desai P. R., Tegtmeyer H. Proposed molecular basis of murine tumor cell-hepatocyte interaction. J Biol Chem. 1983 May 10;258(9):5702–5706. [PubMed] [Google Scholar]
  18. Springer G. F., Desai P. R., Tegtmeyer H., Carlstedt S. C., Scanlon E. F. T/Tn antigen vaccine is effective and safe in preventing recurrence of advanced human breast carcinoma. Cancer Biother. 1994 Spring;9(1):7–15. doi: 10.1089/cbr.1994.9.7. [DOI] [PubMed] [Google Scholar]
  19. Springer G. F., Desai P. R., Tegtmeyer H., Spencer B. D., Scanlon E. F. Pancarcinoma T/Tn antigen detects human carcinoma long before biopsy does and its vaccine prevents breast carcinoma recurrence. Ann N Y Acad Sci. 1993 Aug 12;690:355–357. doi: 10.1111/j.1749-6632.1993.tb44029.x. [DOI] [PubMed] [Google Scholar]
  20. Springer G. F., Desai P. R., Wise W., Carlstedt S. C., Tegtmeyer H., Stein R., Scanlon E. F. Pancarcinoma T and Tn epitopes: autoimmunogens and diagnostic markers that reveal incipient carcinomas and help establish prognosis. Immunol Ser. 1990;53:587–612. [PubMed] [Google Scholar]
  21. Springer G. F. T and Tn, general carcinoma autoantigens. Science. 1984 Jun 15;224(4654):1198–1206. doi: 10.1126/science.6729450. [DOI] [PubMed] [Google Scholar]
  22. Thatcher N., Hashmi K., Chang J., Swindell R., Crowther D. Anti-T antibody in malignant melanoma patients. Influence of response and survival following chemotherapy--changes in serum levels following C parvum, BCG immunization. Cancer. 1980 Sep 15;46(6):1378–1382. doi: 10.1002/1097-0142(19800915)46:6<1378::aid-cncr2820460615>3.0.co;2-n. [DOI] [PubMed] [Google Scholar]
  23. Van Heyningen W. E., Carpenter C. C., Pierce N. F., Greenough W. B., 3rd Deactivation of cholera toxin by ganglioside. J Infect Dis. 1971 Oct;124(4):415–418. doi: 10.1093/infdis/124.4.415. [DOI] [PubMed] [Google Scholar]
  24. Vos G. H., Brain P. Heterophile antibodies, immunoglobulin levels, and the evaluation on anti-T activity in cancer patients and controls. S Afr Med J. 1981 Jul 25;60(4):133–136. [PubMed] [Google Scholar]

Articles from Protein Science : A Publication of the Protein Society are provided here courtesy of The Protein Society

RESOURCES