title: The Escherichia coli heat-stable enterotoxin is a long-lived superagonist of guanylin.
abstract: The mechanism by which bacterial heat-stable enterotoxins (ST I STA) cause diarrhea in humans and animals has been linked to the activation of an intestinal membrane-bound guanylate cyclase. Guanylin, a recently discovered rat intestinal peptide, is homologous in structure to ST I and can activate guanylate cyclase present on the human colonic carcinoma cell line T84. To directly test the mechanistic association of guanylate cyclase activation with diarrhea, we synthesized guanylin and a guanylin analog termed N9P10 guanylin and compared their biological activities with those of a synthetic ST I analog, termed ST Ib(6-18). We report that guanylin is able to inhibit the binding of a radiolabeled ST I analog to rat intestinal cells but causes diarrhea in infant mice only at doses at least 4 orders of magnitude higher than that of ST Ib(6-18). In contrast, N9P10 guanylin was enterotoxic in mice at much lower doses than guanylin but proved to be a weaker inhibitor of radiolabeled ST I than guanylin in the receptor binding assay. The pattern of guanylate cyclase activation observed for ST Ib(6-18) and the two guanylin analogs parallels the results observed in the receptor binding assay rather than those observed in the diarrheal assay. Treatment of guanylin with chymotrypsin or lumenal fluid derived from newborn mouse intestines resulted in a rapid loss of binding activity. Together, these results suggest that ST I enterotoxins may represent a class of long-lived superagonists of guanylin.
authors.affiliation: Department of Medical Biophysics, University of Toronto, Ontario, Canada.
authors.forename: B W
authors.initials: BW
authors.lastname: Carpick
authors.forename: J
authors.initials: J
authors.lastname: Gariépy
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000833:Animals, Suckling; D001427:Bacterial Toxins / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D003967:Diarrhea / Q000139:chemically induced; D004768:Enterotoxins / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D004789:Enzyme Activation / Q000187:drug effects; D029968:Escherichia coli Proteins; D005768:Gastrointestinal Hormones; D006162:Guanylate Cyclase / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism; D007421:Intestine, Small / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D045265:Natriuretic Peptides; D010455:Peptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D000074261:Receptors, Enterotoxin; D050705:Receptors, Guanylate Cyclase-Coupled; D018000:Receptors, Peptide / Q000378:metabolism
chemical_list: D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D005768:Gastrointestinal Hormones; D045265:Natriuretic Peptides; D010455:Peptides; D018000:Receptors, Peptide; C034151:heat stable toxin (E coli); C072685:guanylin; D006162:Guanylate Cyclase; D000074261:Receptors, Enterotoxin; D050705:Receptors, Guanylate Cyclase-Coupled