Radiolabeled toxin for studying binding of cholera toxin and toxoids to intestinal mucosal receptor sites

Johnny Peterson, W. F. Verwey

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Recent evidence suggests that the mode of action of the cholera toxin involves at least 2 steps, which include binding to a membrane receptor and stimulation of the membrane bound enzyme adenyl cyclase. A sensitive in vitro technique for studying toxin binding to tissue receptor sites (guinea pig intestinal cells) is described and appears to have considerable potential usefulness in the study of toxin interaction with mucosal tissue. Highly purified 125I labeled cholera toxin was found to bind specifically to insoluble mucosal homogenates from the small intestines of normal adult guinea pigs. Binding of labeled toxin could be inhibited by nanogram levels of both cholera toxin and choleragenoid (a spontaneously formed toxoid), but not by heterologous rabbit serum proteins. Artificial cholera toxoids prepared by treatment of purified toxin with formaldehyde or glutaraldehyde failed to inhibit radiolabeled toxin binding to mucosal homogenates. The data indicate that cholera toxin binding is highly specific for the toxin and is not inhibited by heterologous proteins. In contrast, artificial toxoids, currently under investigation as potential human vaccines, appear to have lost their affinity for the tissue receptor.

Original languageEnglish (US)
Pages (from-to)1187-1191
Number of pages5
JournalProceedings of the Society for Experimental Biology and Medicine
Volume145
Issue number4
StatePublished - 1974

Fingerprint

Cholera Toxin
Toxoids
Guinea Pigs
Tissue
Membranes
Glutaral
Adenylyl Cyclases
Formaldehyde
Small Intestine
Blood Proteins
Mucous Membrane
Vaccines
cholera toxoid
Rabbits
Enzymes
Proteins

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

@article{1aca6baee7cd4d408588a37e4e20fadf,
title = "Radiolabeled toxin for studying binding of cholera toxin and toxoids to intestinal mucosal receptor sites",
abstract = "Recent evidence suggests that the mode of action of the cholera toxin involves at least 2 steps, which include binding to a membrane receptor and stimulation of the membrane bound enzyme adenyl cyclase. A sensitive in vitro technique for studying toxin binding to tissue receptor sites (guinea pig intestinal cells) is described and appears to have considerable potential usefulness in the study of toxin interaction with mucosal tissue. Highly purified 125I labeled cholera toxin was found to bind specifically to insoluble mucosal homogenates from the small intestines of normal adult guinea pigs. Binding of labeled toxin could be inhibited by nanogram levels of both cholera toxin and choleragenoid (a spontaneously formed toxoid), but not by heterologous rabbit serum proteins. Artificial cholera toxoids prepared by treatment of purified toxin with formaldehyde or glutaraldehyde failed to inhibit radiolabeled toxin binding to mucosal homogenates. The data indicate that cholera toxin binding is highly specific for the toxin and is not inhibited by heterologous proteins. In contrast, artificial toxoids, currently under investigation as potential human vaccines, appear to have lost their affinity for the tissue receptor.",
author = "Johnny Peterson and Verwey, {W. F.}",
year = "1974",
language = "English (US)",
volume = "145",
pages = "1187--1191",
journal = "Experimental Biology and Medicine",
issn = "1535-3702",
publisher = "SAGE Publications Ltd",
number = "4",

}

TY - JOUR

T1 - Radiolabeled toxin for studying binding of cholera toxin and toxoids to intestinal mucosal receptor sites

AU - Peterson, Johnny

AU - Verwey, W. F.

PY - 1974

Y1 - 1974

N2 - Recent evidence suggests that the mode of action of the cholera toxin involves at least 2 steps, which include binding to a membrane receptor and stimulation of the membrane bound enzyme adenyl cyclase. A sensitive in vitro technique for studying toxin binding to tissue receptor sites (guinea pig intestinal cells) is described and appears to have considerable potential usefulness in the study of toxin interaction with mucosal tissue. Highly purified 125I labeled cholera toxin was found to bind specifically to insoluble mucosal homogenates from the small intestines of normal adult guinea pigs. Binding of labeled toxin could be inhibited by nanogram levels of both cholera toxin and choleragenoid (a spontaneously formed toxoid), but not by heterologous rabbit serum proteins. Artificial cholera toxoids prepared by treatment of purified toxin with formaldehyde or glutaraldehyde failed to inhibit radiolabeled toxin binding to mucosal homogenates. The data indicate that cholera toxin binding is highly specific for the toxin and is not inhibited by heterologous proteins. In contrast, artificial toxoids, currently under investigation as potential human vaccines, appear to have lost their affinity for the tissue receptor.

AB - Recent evidence suggests that the mode of action of the cholera toxin involves at least 2 steps, which include binding to a membrane receptor and stimulation of the membrane bound enzyme adenyl cyclase. A sensitive in vitro technique for studying toxin binding to tissue receptor sites (guinea pig intestinal cells) is described and appears to have considerable potential usefulness in the study of toxin interaction with mucosal tissue. Highly purified 125I labeled cholera toxin was found to bind specifically to insoluble mucosal homogenates from the small intestines of normal adult guinea pigs. Binding of labeled toxin could be inhibited by nanogram levels of both cholera toxin and choleragenoid (a spontaneously formed toxoid), but not by heterologous rabbit serum proteins. Artificial cholera toxoids prepared by treatment of purified toxin with formaldehyde or glutaraldehyde failed to inhibit radiolabeled toxin binding to mucosal homogenates. The data indicate that cholera toxin binding is highly specific for the toxin and is not inhibited by heterologous proteins. In contrast, artificial toxoids, currently under investigation as potential human vaccines, appear to have lost their affinity for the tissue receptor.

UR - http://www.scopus.com/inward/record.url?scp=0016292068&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0016292068&partnerID=8YFLogxK

M3 - Article

C2 - 4827729

AN - SCOPUS:0016292068

VL - 145

SP - 1187

EP - 1191

JO - Experimental Biology and Medicine

JF - Experimental Biology and Medicine

SN - 1535-3702

IS - 4

ER -