Isolation and identification of AIDA-I receptors in porcine intestinal mucus.
Porcine AIDA-I positive Escherichia coli causes diarrhea in neonatal piglets and AIDA-I adhesin is an important virulence factor involved in intestinal colonization with biofilm formation. This biofilm consists of AIDA-I(+)E. coli bacteria stratified within mucus layers covering the intestinal mucosa. Based on the intimate interaction between AIDA-I(+)E. coli and mucus within the intestinal biofilm, we hypothesized that porcine intestinal mucus contains receptor(s) for AIDA-I adhesin. Since porcine AIDA-I receptors have not been identified, we employed affinity chromatography and in vitro adhesion assays to investigate AIDA-I binding proteins in porcine intestinal mucus that might serve as receptors for attachment of AIDA-I positive E. coli. We demonstrated that porcine mucus contains 65 and 120kDa proteins (p65 and p120) that bind with high affinity to purified AIDA-I adhesin and that AIDA-I positive E. coli binds to these proteins with higher affinity than do AIDA-I negative mutant. The identity of p65 was not determined based on LC-MS/MS data, whereas p120 was matched to two nuclear proteins (namely, DNA damage binding protein and splicing factor 3b) and one cytoplasmic protein, which is an IgG Fc binding protein. Based on similar amino acid homology, molecular weight, structural similarity to mucin and reported evidence of being secreted by goblet cells into the intestinal lumen, we think that the IgG Fc binding protein is most likely candidate to serve as a potential receptor in intestinal mucus for AIDA-I adhesin.
Fang
Y
Y
Department of Veterinary Pathology, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, SK, Canada S7N 5B4.
Ngeleka
M
M
Middleton
D M
DM
Simko
E
E
D018830:Adhesins, Escherichia coli / Q000502:physiology*; D000818:Animals; D000831:Animals, Newborn; D001422:Bacterial Adhesion / Q000502:physiology; D018441:Biofilms / Q000254:growth & development*; D015153:Blotting, Western / Q000662:veterinary; D002846:Chromatography, Affinity / Q000662:veterinary; D002853:Chromatography, Liquid / Q000662:veterinary; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004926:Escherichia coli / Q000254:growth & development / Q000472:pathogenicity*; D004927:Escherichia coli Infections / Q000382:microbiology / Q000662:veterinary*; D007413:Intestinal Mucosa / Q000382:microbiology; D013552:Swine; D013553:Swine Diseases / Q000382:microbiology*; D053719:Tandem Mass Spectrometry / Q000662:veterinary; D037521:Virulence Factors
C075559:AIDA-I protein, E coli; D018830:Adhesins, Escherichia coli; D037521:Virulence Factors