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Infection and immunity1984; 46(2); 578-584; doi: 10.1128/iai.46.2.578-584.1984

Isolation and structural characterization of the equine erythrocyte receptor for enterotoxigenic Escherichia coli K99 fimbrial adhesin.

Abstract: The erythrocyte receptor for Escherichia coli K99 fimbrial adhesin was isolated from equine erythrocytes and characterized as Neu5Gc-alpha(2----3)-Galp-beta(1----4)-GLcp-beta(1----1)-Ceramide. This glycolipid acted as the receptor for K99 by four different experimental approaches: inhibition of equine erythrocyte hemagglutination by preincubation of K99-positive bacteria or purified K99 fimbriae with the isolated glycolipid; inhibition of attachment of K99-positive bacteria to porcine intestinal epithelial cells in the presence of the isolated glycolipid; induction of binding of K99-positive bacteria or purified K99 fimbriae to normally unreactive guinea pig erythrocytes by coating these cells with the isolated glycolipid; and isolation of the receptor by affinity chromatography with K99 coupled to CNBr-activated Sepharose 4B, indicating a strong interaction between K99 and the isolated glycolipid.
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Publication Date: 1984-11-01 PubMed ID: 6150011PubMed Central: PMC261574DOI: 10.1128/iai.46.2.578-584.1984Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article is about the identification and analysis of the receptor in horse red blood cells that binds to a specific protein found on a type of E. coli, emphasizing its structure and role in bacterial attachment.

Isolating the Receptor

  • The main focus of the paper is the isolation and characterization of the erythrocyte (red blood cell) receptor for Escherichia coli K99 fimbrial adhesin from equine (horse) erythrocytes.
  • This receptor is a glycolipid – a molecule composed of a carbohydrate attached to a lipid – and its complex structure is indicated by the code Neu5Gc-alpha(2—-3)-Galp-beta(1—-4)-GLcp-beta(1—-1)-Ceramide.

Experimental Approaches

  • Four experimental approaches were used to confirm that this glycolipid acts as the receptor for K99.
  • First, the researchers preincubated K99-positive bacteria or purified K99 fimbriae (a structure that allows the bacteria to stick to cells) with the isolated glycolipid, which inhibited the bacteria’s ability to agglutinate (clump together) equine erythrocytes. This suggests that the receptor binds to the bacteria and prevents them from interacting with other cells.
  • Second, they showed that the K99-positive bacteria could not attach to porcine (pig) intestinal epithelial cells when the isolated glycolipid was present. This also indicates that the bacteria bind to the glycolipid instead of the porcine cells.
  • Third, the researchers found that coating unreactive guinea pig erythrocytes with the isolated glycolipid induced binding of the K99-positive bacteria or purified K99 fimbriae. This demonstrates that the bacterial protein specifically interacts with the glycolipid.
  • The final approach involved using affinity chromatography (a method for separating biochemical mixtures) with K99 coupled to CNBr-activated Sepharose 4B. The fact that the receptor glycolipid was isolated in this process indicates a strong interaction between the K99 protein and the glycolipid.

Conclusion of Research

  • In conclusion, the research establishes a solid interaction between the K99 protein found in E. coli and an isolated glycolipid receptor from equine erythrocytes. The study reveals that the bacteria’s ability to adhere to cells is significantly influenced by this interaction. This deeper understanding might contribute to better therapies or preventive measures against E. coli infections in the future.

Cite This Article

APA
Smit H, Gaastra W, Kamerling JP, Vliegenthart JF, de Graaf FK. (1984). Isolation and structural characterization of the equine erythrocyte receptor for enterotoxigenic Escherichia coli K99 fimbrial adhesin. Infect Immun, 46(2), 578-584. https://doi.org/10.1128/iai.46.2.578-584.1984

Publication

Infection and immunity
ISSN: 0019-9567
NlmUniqueID: 0246127
Country: United States
Language: English
Volume: 46
Issue: 2
Pages: 578-584

Researcher Affiliations

Smit, H
    Gaastra, W
      Kamerling, J P
        Vliegenthart, J F
          de Graaf, F K

            MeSH Terms

            • Adhesins, Escherichia coli
            • Adhesiveness
            • Animals
            • Bacterial Proteins / immunology
            • Carbohydrate Sequence
            • Erythrocyte Membrane / analysis
            • Escherichia coli / immunology
            • Fimbriae, Bacterial / immunology
            • Fimbriae, Bacterial / metabolism
            • Glycolipids / isolation & purification
            • Guanylate Cyclase
            • Guinea Pigs
            • Hemagglutination
            • Horses / blood
            • Intestinal Mucosa / immunology
            • Magnetic Resonance Spectroscopy
            • Receptors, Enterotoxin
            • Receptors, Guanylate Cyclase-Coupled
            • Receptors, Immunologic / isolation & purification
            • Receptors, Immunologic / metabolism
            • Receptors, Peptide

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