Campylobacter jejuni FlpA binds fibronectin and is required for maximal host cell adherence

Michael E Konkel; Charles L Larson; Rebecca C Flanagan

doi:10.1128/JB.00969-09

Back

Campylobacter jejuni FlpA binds fibronectin and is required for maximal host cell adherence

Journal article

Open access

Peer reviewed

Campylobacter jejuni FlpA binds fibronectin and is required for maximal host cell adherence

Michael E Konkel, Charles L Larson and Rebecca C Flanagan

Journal of bacteriology, Vol.192(1), pp.68-76

01/2010

DOI: https://doi.org/10.1128/JB.00969-09

Handle:

https://hdl.handle.net/2376/109730

PMCID: PMC2798237

PMID: 19880595

Files and links (1)

url

https://doi.org/10.1128/JB.00969-09View

Published (Version of record) Open

Abstract

Cell Line

Rabbits

Transcription Factors - physiology

Enzyme-Linked Immunosorbent Assay

Electrophoresis, Polyacrylamide Gel

Humans

Bacterial Proteins - genetics

Transcription Factors - genetics

Bacterial Proteins - physiology

Genetic Complementation Test

Fibronectins - metabolism

Transcription Factors - metabolism

Animals

Bacterial Adhesion - physiology

Bacterial Adhesion - genetics

Campylobacter jejuni - metabolism

Protein Binding

Bacterial Proteins - metabolism

Female

Fibronectins - genetics

Fluorescent Antibody Technique, Indirect

Campylobacter jejuni - genetics

Campylobacter jejuni is one of the most frequent bacterial causes of food-borne gastrointestinal disease in developed countries. Previous work indicates that the binding of C. jejuni to human intestinal cells is crucial for host colonization and disease. Fibronectin (Fn), a major constituent of the extracellular matrix, is a approximately 250-kDa glycoprotein present at regions of cell-to-cell contact in the intestinal epithelium. Fn is composed of three types of repeating units: type I (approximately 45 amino acids), type II (approximately 60 amino acids), and type III (approximately 90 amino acids). The deduced amino acid sequence of C. jejuni flpA (Cj1279c) contains at least three Fn type III domains. Based on the presence of the Fn type III domains, we hypothesized that FlpA contributes to the binding of C. jejuni to human INT 407 epithelial cells and Fn. We assessed the contribution of FlpA in C. jejuni binding to host cells by in vitro adherence assays with a C. jejuni wild-type strain and a C. jejuni flpA mutant and binding of purified FlpA protein to Fn by enzyme-linked immunosorbent assay (ELISA). Adherence assays revealed the binding of the C. jejuni flpA mutant to INT 407 epithelial cells was significantly reduced compared with that for a wild-type strain. In addition, rabbit polyclonal serum generated against FlpA blocked C. jejuni adherence to INT 407 cells in a concentration-dependent manner. Binding of FlpA to Fn was found to be dose dependent and saturable by ELISA, demonstrating the specificity of the interaction. Based on these data, we conclude that FlpA mediates C. jejuni attachment to host epithelial cells via Fn binding.

Metrics

13 Record Views

Details

Title: Campylobacter jejuni FlpA binds fibronectin and is required for maximal host cell adherence
Creators: Michael E Konkel - Department of Microbiology, School of Molecular Biosciences, Washington State University, Pullman, Washington, USA. konkel@wsu.edu
Charles L Larson
Rebecca C Flanagan
Publication Details: Journal of bacteriology, Vol.192(1), pp.68-76
Academic Unit: Molecular Biosciences, School of
Publisher: United States
Identifiers: 99900547102401842
Language: English
Resource Type: Journal article