Chmielewski, Markus and Kuehle, Johannes and Chrobok, Danuta and Riet, Nicole and Hallek, Michael and Abken, Hinrich (2019) FimH-based display of functional eukaryotic proteins on bacteria surfaces. SCIENTIFIC REPORTS, 9: UNSP 8410. ISSN 2045-2322,
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The demand for recombinant proteins for analytic and therapeutic purposes is increasing; however, most currently used bacterial production systems accumulate the recombinant proteins in the intracellular space, which requires denaturating procedures for harvesting and functional testing. We here present a novel FimH-based expression system that enables display of fully functional eukaryotic proteins while preventing technical difficulties in translocating, folding, stabilizing and isolating the displayed proteins. As examples, Gaussia Luciferase (GLuc), epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha) and epiregulin (EPRG) were expressed as FimH fusion proteins on the surface of E. coli bacteria. The fusion proteins were functionally active and could be released from the bacterial surface by specific proteolytic cleavage into the culture supernatant allowing harvesting of the produced proteins. EGFR ligands, produced as FimH fusion proteins and released by proteolytic cleavage, bound to the EGF receptor (EGFR) on cancer cells inducing EGFR phosphorylation. In another application of the technology, GLuc-FimH expressed on the surface of bacteria was used to track tumor-infiltrating bacteria by bioluminescence imaging upon application to mice, thereby visualizing the colonization of transplanted tumors. The examples indicate that the FimH-fusion protein technology can be used in various applications that require functionally active proteins to be displayed on bacterial surfaces or released into the culture supernatant.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | OUTER-MEMBRANE; ESCHERICHIA-COLI; CELL-SURFACE; BETA-LACTAMASE; AUTODISPLAY; MECHANISM; FIMBRIAE; ADHESIN; YIELDS; PILUS; |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Immunologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 08 Apr 2020 09:55 |
| Last Modified: | 08 Apr 2020 09:55 |
| URI: | https://pred.uni-regensburg.de/id/eprint/26842 |
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