Design of a Photoreactive Analogue of the Escherichia Coli Heat-stable Enterotoxin STIb: Use in Identifying Its Receptor on Rat Brush Border Membranes

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1986 Jan 1

PMID 3510436

Citations 8

Authors

J Gariepy

G K Schoolnik

Affiliations

Soon will be listed here.

Abstract

The Escherichia coli heat-stable enterotoxin, STIb was prepared by solid-phase peptide synthesis and purified to homogeneity by high-pressure liquid chromatography. This analogue was iodinated and shown to bind specifically to rat intestinal membranes. The radiolabeled peptide was derivatized at the amino terminus with the photoreactive heterobifunctional crosslinking agent N-hydroxysuccinimidyl p-benzoylbenzoate. This photoreactive probe also exhibited binding specificity. It was mixed with rat intestinal brush border membranes and photolyzed in the presence or absence of excess unlabeled STIb. Polyacrylamide gel electrophoresis performed in the presence of sodium dodecyl sulfate and 2-mercaptoethanol indicated that the peptide probe was crosslinked specifically to two molecular species of 57 and 75 kDa. One or both of these molecules appear to constitute the enterotoxin receptor or to be in close proximity to it.

Citing Articles

Interaction of Escherichia coli heat-stable enterotoxin B with cultured human intestinal epithelial cells.

Chao K, Dreyfus L Infect Immun. 1997; 65(8):3209-17.

PMID: 9234777 PMC: 175454. DOI: 10.1128/iai.65.8.3209-3217.1997.

The Escherichia coli heat-stable enterotoxin is a long-lived superagonist of guanylin.

Carpick B, Gariepy J Infect Immun. 1993; 61(11):4710-5.

PMID: 8104900 PMC: 281225. DOI: 10.1128/iai.61.11.4710-4715.1993.

Substitutions of cysteine residues of Escherichia coli heat-stable enterotoxin by oligonucleotide-directed mutagenesis.

Okamoto K, Yukitake J, Kawamoto Y, Miyama A Infect Immun. 1987; 55(9):2121-5.

PMID: 3305364 PMC: 260666. DOI: 10.1128/iai.55.9.2121-2125.1987.

Reduction of enterotoxic activity of Escherichia coli heat-stable enterotoxin by substitution for an asparagine residue.

Okamoto K, Yukitake J, Miyama A Infect Immun. 1988; 56(8):2144-8.

PMID: 3294186 PMC: 259536. DOI: 10.1128/iai.56.8.2144-2148.1988.

Importance of disulfide bridges in the structure and activity of Escherichia coli enterotoxin ST1b.

Gariepy J, Judd A, Schoolnik G Proc Natl Acad Sci U S A. 1987; 84(24):8907-11.

PMID: 2827159 PMC: 299660. DOI: 10.1073/pnas.84.24.8907.

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