Uropathogenic Escherichia coli is a leading cause of urinary tract infections and bacterial prostatitis, the most common UTI complication in men. The initial stages of a successful infection involve bacterial adhesion to host cells through specialized adhesins. FimH, a protein located at the tip of type 1 pili, plays a crucial role as the main mediator for UPEC binding to bladder cells. While the host partners of FimH in the bladder are well-established, the interactions between FimH and prostate cells remain elusive. Consequently, the overarching goal is to enhance comprehension of the initial steps in prostate infection by investigating the interaction of FimH with prostate proteins. To achieve this, a recombinant FimH was constructed and expressed in an inducible expression vector, and an immunofluorescence staining assay was performed which demonstrated distinctive binding patterns in prostate cells compared to the bladder cell line. A Far Western overlay assay, revealed six distinct protein bands in human prostate cells and two in mouse prostate cells, indicating different potential protein partners. These interactions were examined under native conditions by establishing and optimizing a co-immunoprecipitation assay with cell proteins derived from both human and mouse prostates, with the 5637 cell line serving as a positive control. In summary, this study reveals striking differences between FimH binding to prostate and bladder cells, emphasizing the importance of FimH in adhesion and the need for further exploration of FimH interaction with prostate cells.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:his-23752 |
| Date | January 2023 |
| Creators | Joshi, Amruta Ananta |
| Publisher | Högskolan i Skövde, Institutionen för biovetenskap |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Student thesis, info:eu-repo/semantics/bachelorThesis, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
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