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Discussion
Summary of the main achievements
- We designed the self-assembling polyhedra based on combinations of coiled-coil-forming segment as the polypeptide equivalent of DNA origami producing structures that do not exist in nature. We analyzed the topology of possible combinations of coiled-coil-forming segments and for the first time demonstrated the feasibility of this approach on a polypeptide composed of three designed coiled-coil-forming segments. This polypeptide can by slow annealing assemble a box or two-dimensional polypeptide lattice, depending on its concentration. Formation of nanoscale assembly and polypeptide lattice was confirmed by AFM and TEM.
- We prepared polypeptide assemblies, based on gyrase B fragment, which can be assembled and disassembled by the addition of different compounds, which has particularly useful properties for drug delivery.
- We proposed an approach for biomaterial design that could be crosslinked through coiled-coil interactions, which will allow introducing additional functional polypeptides that provide different functions to the cell-growth matrix, such as antimicrobial activity (LL-37 peptide), cell differentiation (NGF), growth factors etc, with an almost unlimited number of potential combinations.
- We prepared polypeptide network based on self-assembly of a polypeptide that contains a p53 tetramerization domain and a coiled coil domain. Real world application such as ultrafiltration was demonstrated on self-assembled polypeptide membrane for the removal of viruses.
- We introduced a BioBrick standard variant ([http://dspace.mit.edu/bitstream/handle/1721.1/46705/BBFRFC37.pdf?sequence=1 BBF RFC 37]) that allows modification of the length of the linker between polypeptide building blocks and prepared over 100 new BioBricks comprising different coiled-coil combinations, oligomerizing polypeptide domains and functional domains that are dedicated to the manufacturing of polypeptide nanoassemblies.
- We introduced a procedure that streamlines and unifies manufacturing of polypeptides which results, in principle independently of their sequence, in high yield polypeptide production in bacteria regardless of their toxicity, sensitivity to proteolysis and minimizes the purification steps.
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