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PMID: 8698855 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Rabbit sucrase-isomaltase contains a functional intestinal receptor for Clostridium difficile toxin A.

The Journal of clinical investigation ·Vol. 98 ·No. 3 ·1996-08-01 ·Pages 641-9

Pothoulakis C, Gilbert RJ, Cladaras C, Castagliuolo I, Semenza G, Hitti Y, Montcrief JS, Linevsky J, Kelly CP, Nikulasson S, Desai HP, Wilkins TD, LaMont JT

Abstract

The intestinal effects of Clostridium difficile toxin A are inidated by toxin binding to luminal enterocyte receptors. We reported previously that the rabbit ileal brush border (BB) receptor is a glycoprotein with an alpha-d-galactose containing trisaccharide in the toxin-binding domain (1991. J. Clin. Invest. 88:119-125). In this study we characterized the rabbit ileal BB receptor for this toxin. Purified toxin receptor peptides of 19 and 24 amino acids showed 100% homology with rabbit sucrase-isomaltase (SI). Guinea pig receptor antiserum reacted in Western blots with rabbit SI and with the purified toxin receptor. Antireceptor IgG blocked in vitro binding of toxin A to rabbit ileal villus cell BB. Furthermore, anti-SI IgG inhibited toxin A-induced secretion (by 78.1%, P < 0.01), intestinal permeability (by 80.8%, P < 0.01), and histologic injury (P < 0.01) in rabbit ileal loops in vivo. Chinese hamster ovary cells transfected with SI cDNA showed increased intracellular calcium increase in response to native toxin (holotoxin) or to a recombinant 873-amino acid peptide representing the receptor binding domain of toxin A. These data suggest that toxin A binds specifically to carbohydrate domains on rabbit ileal SI, and that such binding is relevant to signal transduction mechanisms that mediate in vitro and in vivo toxicity.

MeSH Terms
Amino Acid Sequence Animals Bacterial Toxins Binding Sites CHO Cells Clostridioides difficile/pathogenicity Cricetinae Enterotoxins/metabolism Guinea Pigs Intestine, Small/metabolism Male Molecular Sequence Data Rabbits Sucrase-Isomaltase Complex/metabolism Transfection
Chemicals
Bacterial Toxins Enterotoxins tcdA protein, Clostridium difficile Sucrase-Isomaltase Complex
Authors & Affiliations
13 authors, click to expand affiliations / ORCID
Pothoulakis C
Section of Gastroenterology, Boston University School of Medicine, Massachusetts, USA.
Gilbert R J
Cladaras C
Castagliuolo I
Semenza G
Hitti Y
Montcrief J S
Linevsky J
Kelly C P
Nikulasson S
Desai H P
Wilkins T D
LaMont J T
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Article Info
Journal
The Journal of clinical investigation
Abbr.
J Clin Invest
ISSN
0021-9738
DOI
10.1172/JCI118835
Published
1996-08-01
Pages
641-9
Language
English
Region
United States
NLM ID
7802877
PMCID
PMC507473
Subset
IM
Grants
NIDDK NIH HHS · DK-34583 · United States
NIDDK NIH HHS · DK-47343 · United States
NIDDK NIH HHS · DK-62128 · United States
External Links
PubMed source DOI: 10.1172/JCI118835 PMC Full Text View journal details
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