A relatively high percentage of porcine <i>Escherichia coli</i> isolates from cases associated with neonatal and post-weaning diarrhea are positive for the gene encoding the adhesin involved in diffuse adherence I (AIDA-I). This gene and its corresponding protein were first identified and characterized in <i>E. coli</i> strain 2787 isolated from human infantile diarrhea. Little is known about the role of the AIDA-I protein in pathogenesis of porcine enteric disease caused by AIDA-I positive E. coli and the properties of AIDA-I protein expressed by porcine AIDA-I positive <i>E. coli</i> isolates and its receptors. <p>In this study, we demonstrated that AIDA-I adhesin isolated from porcine AIDA-I positive <i>E. coli</i> PD20 and PD58 is an acidic protein consisting of five isoforms. It has a molecular weight (100 kDa) similar to the AIDA-I adhesin expressed by human AIDA-I positive <i>E. coli</i> strain 2787 and has a relatively high amino acid homology (78-87%) with it. Immunodetection of AIDA-I positive <i>E. coli</i> strains using polyclonal anti-AIDA-I antibodies had relatively low sensitivity and specificity, accordingly these tests are unlikely to be used for regular diagnostic detection. <p>Using affinity chromatography, we isolated from porcine intestinal mucus proteins that bind to purified AIDA-I adhesin. These proteins were separated by one- and two-dimensional electrophoresis and subjected to overlay Western blot with purified AIDA-I adhesin and AIDA-I positive <i>E. coli</i> to demonstrate 65 and 120 kDa (p65 and p120) proteins as AIDA-I binding proteins. The identity of p65 was not determined based on LCMS/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 the most likely candidate to serve as a potential receptor in intestinal mucus for AIDA-I adhesin.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:SSU.etd-04172007-091643 |
| Date | 25 April 2007 |
| Creators | Fang, Yuanmu |
| Contributors | Simko, Elemir, Middleton, Dorothy M. |
| Publisher | University of Saskatchewan |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://library.usask.ca/theses/available/etd-04172007-091643/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Saskatchewan or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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