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Cell Surface-Associated Elongation Factor Tu Mediates the Attachment of Lactobacillus johnsonii NCC533 (La1) to Human Intestinal Cells and Mucins
- Source :
- Infection and Immunity. 72:2160-2169
- Publication Year :
- 2004
- Publisher :
- American Society for Microbiology, 2004.
-
Abstract
- The aim of this work was to identify Lactobacillus johnsonii NCC533 (La1) surface molecules mediating attachment to intestinal epithelial cells and mucins. Incubation of Caco-2 intestinal epithelial cells with an L. johnsonii La1 cell wall extract led to the recognition of elongation factor Tu (EF-Tu) as a novel La1 adhesin-like factor. The presence of EF-Tu at the surface of La1 was confirmed by analysis of purified outer surface protein extract by immunoblotting experiments, by electron microscopy, and by enzyme-linked immunosorbent assays of live bacteria. Furthermore, tandem mass spectrometry analysis proved that EF-TU was expressed at the La1 surface as an intact molecule. Using recombinant La1 EF-Tu protein, we were able to determine that its binding to intestinal cells and to mucins is pH dependent. Competition experiments suggested that EF-Tu has an important role in La1 mucin binding capacity. In addition, immunomodulation studies performed on HT29 cells showed that EF-Tu recombinant protein can induce a proinflammatory response in the presence of soluble CD14. Our in vitro results indicate that EF-Tu, through its binding to the intestinal mucosa, might participate in gut homeostasis.
- Subjects :
- Molecular Sequence Data
Immunology
Peptide Elongation Factor Tu
Microbiology
Bacterial Adhesion
Cell Line
Mice
HT29 Cells
Intestinal mucosa
Cell Wall
Animals
Humans
Amino Acid Sequence
Interleukin 8
Adhesins, Bacterial
Lactobacillus johnsonii
Inflammation
Mice, Inbred BALB C
Cellular Microbiology: Pathogen-Host Cell Molecular Interactions
biology
Interleukin-8
Mucin
Mucins
biology.organism_classification
Recombinant Proteins
In vitro
Cell biology
Intestines
Lactobacillus
Infectious Diseases
Biochemistry
Caco-2
Cell culture
Parasitology
Caco-2 Cells
Subjects
Details
- ISSN :
- 10985522 and 00199567
- Volume :
- 72
- Database :
- OpenAIRE
- Journal :
- Infection and Immunity
- Accession number :
- edsair.doi.dedup.....601503647c3f567410624c37ff907f03