Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1991 Dec;59(12):4318–4323. doi: 10.1128/iai.59.12.4318-4323.1991

Age-dependent changes in affinity-labeled receptors for Escherichia coli heat-stable enterotoxin in the swine intestine.

L C Katwa 1, C D Parker 1, A A White 1
PMCID: PMC259043  PMID: 1682259

Abstract

Intestinal brush border membranes from 1-day-old and 4-week-old (day of weaning) pigs were affinity labeled with an Escherichia coli heat-stable enterotoxin (STa) by cross-linking 125I-STa to receptor proteins with disuccinimidyl suberate. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed that a radioactive protein with a relative molecular weight of 137,000 to 145,000 was present in both age groups. A strongly radioactive protein with an apparent Mr of 90,000 was present in the 1-day-old animals but not in those that were 4 weeks old. The major radioactive protein present in the older pigs had an Mr of 64,000 to 67,000, but this protein was missing or very weakly radioactive in the younger pigs. There was no significant difference between the groups in receptor affinity for STa, although the receptor density in the older animals was marginally significantly greater. STa-stimulated guanylate cyclase activity in membranes from 1-day-old pigs was only one-sixth that in 4-week-old pigs, although the basal and Lubrol PX-stimulated activities were similar.

Full text

PDF
4318

Images in this article

4320Image
on p.4320

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. 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]
  3. Cohen M. B., Guarino A., Shukla R., Giannella R. A. Age-related differences in receptors for Escherichia coli heat-stable enterotoxin in the small and large intestine of children. Gastroenterology. 1988 Feb;94(2):367–373. doi: 10.1016/0016-5085(88)90423-4. [DOI] [PubMed] [Google Scholar]
  4. Cohen M. B., Moyer M. S., Luttrell M., Giannella R. A. The immature rat small intestine exhibits an increased sensitivity and response to Escherichia coli heat-stable enterotoxin. Pediatr Res. 1986 Jun;20(6):555–560. doi: 10.1203/00006450-198606000-00017. [DOI] [PubMed] [Google Scholar]
  5. DeBlasi A., O'Reilly K., Motulsky H. J. Calculating receptor number from binding experiments using same compound as radioligand and competitor. Trends Pharmacol Sci. 1989 Jun;10(6):227–229. doi: 10.1016/0165-6147(89)90266-6. [DOI] [PubMed] [Google Scholar]
  6. ElDeib M. M., Parker C. D., Veum T. L., Zinn G. M., White A. A. Characterization of intestinal brush border guanylate cyclase activation by Escherichia coli heat-stable enterotoxin. Arch Biochem Biophys. 1986 Feb 15;245(1):51–65. doi: 10.1016/0003-9861(86)90189-x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Garbers D. L. Guanylate cyclase, a cell surface receptor. J Biol Chem. 1989 Jun 5;264(16):9103–9106. [PubMed] [Google Scholar]
  10. Gariépy J., Schoolnik G. K. Design of a photoreactive analogue of the Escherichia coli heat-stable enterotoxin STIb: use in identifying its receptor on rat brush border membranes. Proc Natl Acad Sci U S A. 1986 Jan;83(2):483–487. doi: 10.1073/pnas.83.2.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gerke V., Weber K. Isolation and characterization of mammalian villin and fimbrin, the two bundling proteins of the intestinal microvilli. Eur J Cell Biol. 1983 Sep;31(2):249–255. [PubMed] [Google Scholar]
  12. 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]
  13. Greenberg R. N., Guerrant R. L. E. coli heat-stable enterotoxin. Pharmacol Ther. 1981;13(3):507–531. doi: 10.1016/0163-7258(81)90027-9. [DOI] [PubMed] [Google Scholar]
  14. Guarino A., Cohen M. B., Giannella R. A. Small and large intestinal guanylate cyclase activity in children: effect of age and stimulation by Escherichia coli heat-stable enterotoxin. Pediatr Res. 1987 Jun;21(6):551–555. doi: 10.1203/00006450-198706000-00009. [DOI] [PubMed] [Google Scholar]
  15. Guarino A., Cohen M. B., Overmann G., Thompson M. R., Giannella R. A. Binding of E. coli heat-stable enterotoxin to rat intestinal brush borders and to basolateral membranes. Dig Dis Sci. 1987 Sep;32(9):1017–1026. doi: 10.1007/BF01297193. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Ivens K., Gazzano H., O'Hanley P., Waldman S. A. Heterogeneity of intestinal receptors for Escherichia coli heat-stable enterotoxin. Infect Immun. 1990 Jun;58(6):1817–1820. doi: 10.1128/iai.58.6.1817-1820.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kaushal V., Barnes L. D. Effect of zwitterionic buffers on measurement of small masses of protein with bicinchoninic acid. Anal Biochem. 1986 Sep;157(2):291–294. doi: 10.1016/0003-2697(86)90629-9. [DOI] [PubMed] [Google Scholar]
  19. Kuno T., Andresen J. W., Kamisaki Y., Waldman S. A., Chang L. Y., Saheki S., Leitman D. C., Nakane M., Murad F. Co-purification of an atrial natriuretic factor receptor and particulate guanylate cyclase from rat lung. J Biol Chem. 1986 May 5;261(13):5817–5823. [PubMed] [Google Scholar]
  20. Kuno T., Kamisaki Y., Waldman S. A., Gariepy J., Schoolnik G., Murad F. Characterization of the receptor for heat-stable enterotoxin from Escherichia coli in rat intestine. J Biol Chem. 1986 Jan 25;261(3):1470–1476. [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Maack T., Suzuki M., Almeida F. A., Nussenzveig D., Scarborough R. M., McEnroe G. A., Lewicki J. A. Physiological role of silent receptors of atrial natriuretic factor. Science. 1987 Oct 30;238(4827):675–678. doi: 10.1126/science.2823385. [DOI] [PubMed] [Google Scholar]
  23. Mezoff A. G., Jensen N. J., Cohen M. B. Mechanisms of increased susceptibility of immature and weaned pigs to Escherichia coli heat-stable enterotoxin. Pediatr Res. 1991 May;29(5):424–428. doi: 10.1203/00006450-199105010-00003. [DOI] [PubMed] [Google Scholar]
  24. Nakazawa M., Sugimoto C., Isayama Y., Kashiwazaki M. Virulence factors in Escherichia coli isolated from piglets with neonatal and post-weaning diarrhea in Japan. Vet Microbiol. 1987 Apr;13(4):291–300. doi: 10.1016/0378-1135(87)90060-5. [DOI] [PubMed] [Google Scholar]
  25. Rao M. C., Field M. Enterotoxins and ion transport. Biochem Soc Trans. 1984 Apr;12(2):177–180. doi: 10.1042/bst0120177. [DOI] [PubMed] [Google Scholar]
  26. Rao M. C., Guandalini S., Smith P. L., Field M. Mode of action of heat-stable Escherichia coli enterotoxin. Tissue and subcellular specificities and role of cyclic GMP. Biochim Biophys Acta. 1980 Sep 17;632(1):35–46. doi: 10.1016/0304-4165(80)90247-0. [DOI] [PubMed] [Google Scholar]
  27. Schenk D. B., Phelps M. N., Porter J. G., Fuller F., Cordell B., Lewicki J. A. Purification and subunit composition of atrial natriuretic peptide receptor. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1521–1525. doi: 10.1073/pnas.84.6.1521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schulz S., Green C. K., Yuen P. S., Garbers D. L. Guanylyl cyclase is a heat-stable enterotoxin receptor. Cell. 1990 Nov 30;63(5):941–948. doi: 10.1016/0092-8674(90)90497-3. [DOI] [PubMed] [Google Scholar]
  29. Takayanagi R., Inagami T., Snajdar R. M., Imada T., Tamura M., Misono K. S. Two distinct forms of receptors for atrial natriuretic factor in bovine adrenocortical cells. Purification, ligand binding, and peptide mapping. J Biol Chem. 1987 Sep 5;262(25):12104–12113. [PubMed] [Google Scholar]
  30. Thompson M. R., Giannella R. A. Different crosslinking agents identify distinctly different putative Escherichia coli heat-stable enterotoxin rat intestinal cell receptor proteins. J Recept Res. 1990;10(1-2):97–117. doi: 10.3109/10799899009064660. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Waldman S. A., Kuno T., Kamisaki Y., Chang L. Y., Gariepy J., O'Hanley P., Schoolnik G., Murad F. Intestinal receptor for heat-stable enterotoxin of Escherichia coli is tightly coupled to a novel form of particulate guanylate cyclase. Infect Immun. 1986 Jan;51(1):320–326. doi: 10.1128/iai.51.1.320-326.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. White A. A., Karr D. B. Improved two-step method for the assay of adenylate and guanylate cyclase. Anal Biochem. 1978 Apr;85(2):451–460. doi: 10.1016/0003-2697(78)90242-7. [DOI] [PubMed] [Google Scholar]
  34. White A. A., Lad P. J. Detergent interactions and solubilization techniques for membrane guanylyl cyclase. Methods Enzymol. 1991;195:363–373. doi: 10.1016/0076-6879(91)95181-i. [DOI] [PubMed] [Google Scholar]
  35. de Jonge H. R. The mechanism of action of Escherichia coli heat-stable toxin. Biochem Soc Trans. 1984 Apr;12(2):180–184. doi: 10.1042/bst0120180. [DOI] [PubMed] [Google Scholar]
  36. elDeib M. M., Parker C. D., White A. A. Regulation of intestinal mucosa guanylate cyclase by hemin, heme and protoporphyrin IX. Biochim Biophys Acta. 1987 Apr 2;928(1):83–91. doi: 10.1016/0167-4889(87)90088-7. [DOI] [PubMed] [Google Scholar]

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

RESOURCES