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Vonderviszt Ferenc

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Kovács Boglárka
Flagellin: a convenient protein in biosensorics

Aug 30 - szerda

11:15 – 11:30

Bioszenzorika és bio-nanotechnológia

E27

Flagellin: a convenient protein in biosensorics

Boglárka Kovács¹, András Saftics¹, Inna Székács¹, Hajnalka Jankovics², Sandor Kurunczi¹, Ferenc Vonderviszt², Robert Horvath¹

¹Nanobiosensorics Laboratory, Centre for Energy Research, Institute of Technical Physics and Materials Science, Budapest, Hungary

²Bio-nanosystem Laboratory, Research Institute of Biomolecular and Chemical Engineering, University of Pannonia, Veszprém, Hungary

Flagellin is the main building block of bacterial flagellar filaments. Since the filaments are located outside of the cells, cell lysis is not required to purify flagellin. Flagellin consists of 4 domains: D0, D1, D2, and D3. The D0 domain contains amphipathic helical regions with hydrophobic amino acids on one side of the helix. This part of flagellin is disordered in solution, but can be used to anchor the protein on hydrophobic surfaces with the D3 domain pointing towards the solution [1]. The hypervariable D3 domain situated on the filament surface is a largely independent part of the flagellin that can be removed or replaced without disturbing filament formation.

During our work we in-depth characterized the coatings created from flagellin, and influenced the adsorption of the protein with Hofmeister salts [1]. We applied genetically modified high affinity Ni-binding variant as receptor, and demonstrated the unique sensitivity of grating-coupled interferometry [2].

The monolayer of wild-type flagellin mimics the surface of the bacterial flagellar filament, and we hypothesized that oriented flagellin layers have bacteria-repellent properties. To prove this, we studied the adhesion of bacterial E. coli and human cancer cells on oriented wild-type flagellin layers [3,4].

Through genetic modification, specific oligopeptide segments can be also inserted into the D3 domain of flagellin, which can induce cell adhesion through integrin receptors. We studied cancer cell adhesion on the genetically engineered protein layers with label-free optical biosensors [4]. Mammalian cells can recognize flagellin in solution through Toll-like receptors, and the protein can cause innate immune system response. We are studying the above biological mechanisms and its consequences in the adhesion of the flagelljn exposed cells. Our results prove, that flagellin can be used in many ways in creating capture layers in biosensors.

References

[1] Kovacs, B.; Saftics, A.; Biro, A.; Kurunczi, S.; Szalontai, B.; Kakasi, B.; Vonderviszt, F.; Der, A.; Horvath, R. J. Phys. Chem. C 2018, 122 (37), 21375–21386.

[2] Jankovics, H.; Kovacs, B.; Saftics, A.; Gerecsei, T.; Tóth, É.; Szekacs, I.; Vonderviszt, F.; Horvath, R. Sci. Rep. 2020, 1–11.

[3] Kovacs, B.; Patko, D.; Klein, A.; Kakasi, B.; Saftics, A.; Kurunczi, S.; Vonderviszt, F.; Horvath, R. Sensors Actuators B Chem. 2018, 257, 839–845.

[4] Kovacs, B.; Patko, D.; Szekacs, I.; Orgovan, N.; Kurunczi, S.; Sulyok, A.; Khanh, N. Q.; Toth, B.; Vonderviszt, F.; Horvath, R. Acta Biomater. 2016, No. 42, 66–76.