ONLINE - Engineered Repeat Proteins in Nanobiotechnology: the examples of ELPs and CTPRs

CIC nanoGUNE Seminars

Evangelos Georgilis, Self-Assembly Group
nanoGUNE online Webinar
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ONLINE - Engineered Repeat Proteins in Nanobiotechnology: the examples of ELPs and CTPRs ** ** **Engineered Repeat Proteins in Nanobiotechnology: the examples of ELPs and CTPRs** **** Evangelos Georgilis Self-Assembly Group, CIC nanoGUNE **** The use of recombinant DNA technology has led to the development of a wide variety of artificial genes [1,2]. This has enabled the production of protein- based polymers, known as recombinamers. Depending on their architecture, recombinamers can be divided into intrinsically disordered [3] and structured sequences [4]. An important example of disordered sequences are the elastin-like polypeptides (ELPs), which find many applications in tissue engineering and nanomedicine because of their biocompatibility and thermal response [5]. ELPs are comprised of Val-Pro-Gly-Xaa-Gly pentapeptide repeats, where Xaa ≠Pro is the guest residue. These polypeptides aggregate reversibly above a transition temperature (Tt). By incorporating guest residues of different hydrophilicity, it is possible to construct diblock ELPs which thermally assemble into nanoparticles. The first part of this work is focused on my recent PhD project in LCPO (Bordeaux), which had as a goal the development of diblock ELPs for applications as thermoresponsive, self-assembled drug delivery vectors. A series of diblock ELPs were designed, which were composed of an Ile-containing hydrophobic block and a Met-containing block. This system could access nanoparticle formation by chemoselective post-modifications of the Met residue, as was shown by means of turbidimetry and light scattering. On the other hand, the consensus tetratricopeptide repeat proteins (CTPRs) represent a successful design of recombinamers with well-defined architectures [6,7], which may serve to organize different elements in a distinct pattern. CTPRs are comprised of a 34-residue motif folded into a pair of antiparallel α-helices which self-assemble into elongated superhelices. The structural integrity of these proteins renders them interesting candidates for fabrication methods such as electrospinning, where the disruption of protein structure may often occur [8]. The second part of this work is focused on my current work on a joint project between nanoGUNE and biomaGUNE on electrospun fibers of CTPRs. The characterization of these fibers by electron microscopy, Raman spectroscopy and X-ray diffraction indicated the structural integrity of CTPRs, therefore other elements may be precisely organized along the fiber axis. **References** [1] J.C. Rodríguez-Cabello, L. Martín, M. Alonso, F.J. Arias, A.M. Testera, “Recombinamers” as advanced materials for the post-oil age, Polymer (Guildf). 50 (2009) 5159–5169. doi:10.1016/j.polymer.2009.08.032. [2] A. Girotti, A. Fernández-Colino, I.M. López, J.C. Rodríguez-Cabello, F.J. Arias, Elastin-like recombinamers: Biosynthetic strategies and biotechnological applications, Biotechnol. J. 6 (2011) 1174–1186. [3] K.M. Ruff, S. Roberts, A. Chilkoti, R. V. Pappu, Advances in Understanding Stimulus-Responsive Phase Behavior of Intrinsically Disordered Protein Polymers, J. Mol. Biol. 430 (2018) 4619–4635. doi:10.1016/j.jmb.2018.06.031. [4] E.R.G. Main, A.R. Lowe, S.G.J. Mochrie, S.E. Jackson, L. Regan, A recurring theme in protein engineering: The design, stability and folding of repeat proteins, Curr. Opin. Struct. Biol. 15 (2005) 464–471. doi:10.1016/ [5] E. Georgilis, M. Abdelghani, J. Pille, E. Aydinlioglu, J.C.M. van Hest, S. Lecommandoux, E. Garanger, Nanoparticles based on natural, engineered or synthetic proteins and polypeptides for drug delivery applications, Int. J. Pharm. 586 (2020). doi:10.1016/j.ijpharm.2020.119537. [6] S.H. Mejías, B. Sot, R. Guantes, A.L. Cortajarena, Controlled nanometric fibers of self-assembled designed protein scaffolds, Nanoscale. 6 (2014) 10982–10988. doi:10.1039/c4nr01210k. [7] T. Kajander, A.L. Cortajarena, E.R.G. Main, S.G.J. Mochrie, L. Regan, A new folding paradigm for repeat proteins, J. Am. Chem. Soc. 127 (2005) 10188–10190. doi:10.1021/ja0524494. [8] Y. Dror, T. Ziv, V. Makarov, H. Wolf, A. Admon, E. Zussman, Nanofibers made of globular proteins, Biomacromolecules. 9 (2008) 2749–2754. doi:10.1021/bm8005243. \---------- ONLINE seminar: Evangelos Georgilis; Self-Assembly Group - CIC nanoGUNE When: Oct 26, 2020 11:00 AM Madrid **Register in advance for this meeting:** After registering, you will receive a confirmation email containing information about joining the meeting. Thank you very much for your participation!