Pathogenesis of Shigella diarrhea: rabbit intestinal cell microvillus membrane binding site for Shigella toxin

Fuchs, G; Mobassaleh, M; Donohue-Rolfe, A; Montgomery, R K; Grand, R J; Keusch, G T

Title: Pathogenesis of Shigella diarrhea: rabbit intestinal cell microvillus membrane binding site for Shigella toxin

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Abstract

This study examined the binding of purified /sup 125/I-labeled shigella toxin to rabbit jejunal microvillus membranes (MVMs). Toxin binding was concentration dependent, saturable, reversible, and specifically inhibited by unlabeled toxin. The calculated number of toxin molecules bound at 4/sup 0/C was 7.9 X 10(10) (3 X 10(10) to 2 X 10(11))/micrograms of MVM protein or 1.2 X 10(6) per enterocyte. Scatchard analysis showed the binding site to be of a single class with an equilibrium association constant, K, of 4.7 X 10(9) M-1 at 4/sup 0/C. Binding was inversely related to the temperature of incubation. A total of 80% of the labeled toxin binding at 4/sup 0/C dissociated from MVM when the temperature was raised to 37/sup 0/C, but reassociated when the temperature was again brought to 4/sup 0/C. There was no structural or functional change of MVM due to toxin as monitored by electron microscopy or assay of MVM sucrase activity. These studies demonstrate a specific binding site for shigella toxin on rabbit MVMs. The physiological relevance of this receptor remains to be determined.

Authors:: Fuchs, G; Mobassaleh, M; Donohue-Rolfe, A; Montgomery, R K; Grand, R J; Keusch, G T

Publication Date:: Fri Aug 01 00:00:00 EDT 1986

Research Org.:: New England Medical Center, Boston, MA

OSTI Identifier:: 5013845

Resource Type:: Journal Article

Journal Name:: Infect. Immun.; (United States)

Additional Journal Information:: Journal Volume: 2

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; BACTERIAL DISEASES; PATHOGENESIS; TOXINS; RADIORECEPTOR ASSAY; CELL MEMBRANES; CHEMICAL BONDS; DIARRHEA; EPITHELIUM; IN VITRO; INTESTINES; IODINE 125; LABELLED COMPOUNDS; RABBITS; RECEPTORS; SHIGELLA; ANIMAL TISSUES; ANIMALS; ANTIGENS; BACTERIA; BETA DECAY RADIOISOTOPES; BODY; CELL CONSTITUENTS; DAYS LIVING RADIOISOTOPES; DIGESTIVE SYSTEM; DISEASES; ELECTRON CAPTURE RADIOISOTOPES; GASTROINTESTINAL TRACT; INFECTIOUS DISEASES; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; MAMMALS; MATERIALS; MEMBRANE PROTEINS; MEMBRANES; MICROORGANISMS; NUCLEI; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; ORGANS; PROTEINS; RADIOISOTOPES; SYMPTOMS; TISSUES; TOXIC MATERIALS; TRACER TECHNIQUES; VERTEBRATES; 550901* - Pathology- Tracer Techniques

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                    Fuchs, G, Mobassaleh, M, Donohue-Rolfe, A, Montgomery, R K, Grand, R J, and Keusch, G T. Pathogenesis of Shigella diarrhea: rabbit intestinal cell microvillus membrane binding site for Shigella toxin.  United States: N. p., 1986. 
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                    Fuchs, G, Mobassaleh, M, Donohue-Rolfe, A, Montgomery, R K, Grand, R J, & Keusch, G T. Pathogenesis of Shigella diarrhea: rabbit intestinal cell microvillus membrane binding site for Shigella toxin.  United States.

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                    Fuchs, G, Mobassaleh, M, Donohue-Rolfe, A, Montgomery, R K, Grand, R J, and Keusch, G T. 1986.  
        "Pathogenesis of Shigella diarrhea: rabbit intestinal cell microvillus membrane binding site for Shigella toxin".  United States.

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@article{osti_5013845,

  title        = {Pathogenesis of Shigella diarrhea: rabbit intestinal cell microvillus membrane binding site for Shigella toxin},

  author       = {Fuchs, G and Mobassaleh, M and Donohue-Rolfe, A and Montgomery, R K and Grand, R J and Keusch, G T},

  abstractNote = {This study examined the binding of purified /sup 125/I-labeled shigella toxin to rabbit jejunal microvillus membranes (MVMs). Toxin binding was concentration dependent, saturable, reversible, and specifically inhibited by unlabeled toxin. The calculated number of toxin molecules bound at 4/sup 0/C was 7.9 X 10(10) (3 X 10(10) to 2 X 10(11))/micrograms of MVM protein or 1.2 X 10(6) per enterocyte. Scatchard analysis showed the binding site to be of a single class with an equilibrium association constant, K, of 4.7 X 10(9) M-1 at 4/sup 0/C. Binding was inversely related to the temperature of incubation. A total of 80% of the labeled toxin binding at 4/sup 0/C dissociated from MVM when the temperature was raised to 37/sup 0/C, but reassociated when the temperature was again brought to 4/sup 0/C. There was no structural or functional change of MVM due to toxin as monitored by electron microscopy or assay of MVM sucrase activity. These studies demonstrate a specific binding site for shigella toxin on rabbit MVMs. The physiological relevance of this receptor remains to be determined.},

  doi          = {},

  url          = {https://www.osti.gov/biblio/5013845},
  journal      = {Infect. Immun.; (United States)},
number       = ,

  volume       = 2,

  place        = {United States},

  year         = {Fri Aug 01 00:00:00 EDT 1986},

  month        = {Fri Aug 01 00:00:00 EDT 1986}

}

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