Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1987 Feb;55(2):329–334. doi: 10.1128/iai.55.2.329-334.1987

Association and dissociation of Escherichia coli heat-stable enterotoxin from rat brush border membrane receptors.

M B Cohen, M R Thompson, G J Overmann, R A Giannella
PMCID: PMC260330  PMID: 3542830

Abstract

Escherichia coli heat-stable enterotoxin (ST) binds to receptors on rat intestinal cells and brush border membranes (BBM). We devised experiments to examine the reversibility of ST binding. We found that both 125I-labeled ST and native ST were spontaneously dissociable from the BBM receptor. Radiolabeled ST bound to BBM was also dissociated by the addition of avid goat anti-ST antiserum. Furthermore, using a computer program for analysis of ligand binding, we calculated an apparent Ka of 10(8) liters/mol from competitive inhibition and saturation-binding data. This is significantly lower than the value previously reported by others. Our findings, of a lower Ka and a reversible ST-binding process, suggest that a therapeutic strategy of removing bound ST from its receptor or competing with the enterocyte receptor for unbound ST might be successful in terminating ST-induced secretion.

Full text

PDF
329

Selected References

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

  1. Aimoto S., Takao T., Shimonishi Y., Hara S., Takeda T., Takeda Y., Miwatani T. Amino-acid sequence of a heat-stable enterotoxin produced by human enterotoxigenic Escherichia coli. Eur J Biochem. 1982 Dec 15;129(2):257–263. doi: 10.1111/j.1432-1033.1982.tb07047.x. [DOI] [PubMed] [Google Scholar]
  2. Black R. E., Brown K. H., Becker S., Alim A. R., Huq I. Longitudinal studies of infectious diseases and physical growth of children in rural Bangladesh. II. Incidence of diarrhea and association with known pathogens. Am J Epidemiol. 1982 Mar;115(3):315–324. doi: 10.1093/oxfordjournals.aje.a113308. [DOI] [PubMed] [Google Scholar]
  3. Black R. E., Brown K. H., Becker S., Yunus M. Longitudinal studies of infectious diseases and physical growth of children in rural Bangladesh. I. Patterns of morbidity. Am J Epidemiol. 1982 Mar;115(3):305–314. doi: 10.1093/oxfordjournals.aje.a113307. [DOI] [PubMed] [Google Scholar]
  4. Chan S. K., Giannella R. A. Amino acid sequence of heat-stable enterotoxin produced by Escherichia coli pathogenic for man. J Biol Chem. 1981 Aug 10;256(15):7744–7746. [PubMed] [Google Scholar]
  5. Dahlqvist A. Assay of intestinal disaccharidases. Anal Biochem. 1968 Jan;22(1):99–107. doi: 10.1016/0003-2697(68)90263-7. [DOI] [PubMed] [Google Scholar]
  6. Field M., Graf L. H., Jr, Laird W. J., Smith P. L. Heat-stable enterotoxin of Escherichia coli: in vitro effects on guanylate cyclase activity, cyclic GMP concentration, and ion transport in small intestine. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2800–2804. doi: 10.1073/pnas.75.6.2800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Frantz J. C., Jaso-Friedman L., Robertson D. C. Binding of Escherichia coli heat-stable enterotoxin to rat intestinal cells and brush border membranes. Infect Immun. 1984 Feb;43(2):622–630. doi: 10.1128/iai.43.2.622-630.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Giannella R. A., Drake K. W. Effect of purified Escherichia coli heat-stable enterotoxin on intestinal cyclic nucleotide metabolism and fluid secretion. Infect Immun. 1979 Apr;24(1):19–23. doi: 10.1128/iai.24.1.19-23.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Giannella R. A., Drake K. W., Luttrell M. Development of a radioimmunoassay for Escherichia coli heat-stable enterotoxin: comparison with the suckling mouse bioassay. Infect Immun. 1981 Jul;33(1):186–192. doi: 10.1128/iai.33.1.186-192.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Giannella R. A., Luttrell M., Thompson M. Binding of Escherichia coli heat-stable enterotoxin to receptors on rat intestinal cells. Am J Physiol. 1983 Oct;245(4):G492–G498. doi: 10.1152/ajpgi.1983.245.4.G492. [DOI] [PubMed] [Google Scholar]
  11. Guerrant R. L., Hughes J. M., Chang B., Robertson D. C., Murad F. Activation of intestinal guanylate cyclase by heat-stable enterotoxin of Escherichia coli: studies of tissue specificity, potential receptors, and intermediates. J Infect Dis. 1980 Aug;142(2):220–228. doi: 10.1093/infdis/142.2.220. [DOI] [PubMed] [Google Scholar]
  12. Hopfer U., Crowe T. D., Tandler B. Purification of brush border membrane by thiocyanate treatment. Anal Biochem. 1983 Jun;131(2):447–452. doi: 10.1016/0003-2697(83)90197-5. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
  15. Takao T., Shimonishi Y., Kobayashi M., Nishimura O., Arita M., Takeda T., Honda T., Miwatani T. Amino acid sequence of heat-stable enterotoxin produced by Vibrio cholerae non-01. FEBS Lett. 1985 Dec 2;193(2):250–254. doi: 10.1016/0014-5793(85)80163-0. [DOI] [PubMed] [Google Scholar]
  16. Thompson M. R., Brandwein H., LaBine-Racke M., Giannella R. A. Simple and reliable enzyme-linked immunosorbent assay with monoclonal antibodies for detection of Escherichia coli heat-stable enterotoxins. J Clin Microbiol. 1984 Jul;20(1):59–64. doi: 10.1128/jcm.20.1.59-64.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Thompson M. R., Luttrell M., Overmann G., Giannella R. A. Biological and immunological characteristics of 125I-4Tyr and -18Tyr Escherichia coli heat-stable enterotoxin species purified by high-performance liquid chromatography. Anal Biochem. 1985 Jul;148(1):26–36. doi: 10.1016/0003-2697(85)90623-2. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES