IgA1 (immunoglobulin A1) functions as an immune molecule, against microbial invaders and other pathogens, on the surface of animal and human cells, especially on the mucosal membrane. To impair the function of immunoglobulin the human immune system some bacteria secrete site-specific IgA1 proteases to cleave IgA1 at hinge region. The protease has therefore been implicated as a putative virulence factor. Although the alignment of the DNA for the proteases predicted the IgA1 protease domain and catalytic site, lack of experimental evidence hindered in both structural and functional enzymic studies. In order to elucidate and understand the exact activity, this study aims at to analyze catalytic site of IgA1 protease from H. influenza, and thus PCR was used to amplify the DNA for putative reactive site domain (1-885) and protease domain (1-2373), respectively, were cloned in pGEM-T and transferred to the expression vector, pTrcHis-A for production of the recombinant proteins. Analysis of the recombinant proteins shows that the Human IgA1 heavy chain hydrolyzed by the protease domain recombinant protein but putative reactive site. This demonstrates that protease domain is enough for protease activity. The putative reactive site domain alone is incapable in hydrolyzing Human IgA1.
| Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0717108-145502 |
| Date | 17 July 2008 |
| Creators | Lin, Hsien-chang |
| Contributors | Chen-Fu Shaw, Shi-ping He, Chan-Shing Lin |
| Publisher | NSYSU |
| Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
| Language | Cholon |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0717108-145502 |
| Rights | not_available, Copyright information available at source archive |
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