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Regulated assembly Idea Approach.html
From 2009.igem.org
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Summary
Folded protein domains that form oligomers with same or other protein domains could be used to form complex nanostructures. We prepared a new type of polypeptide building blocks composed of dimerization and trimerization domains of proteins, connected by a short flexible peptide linker. This combination should be able to form a polypeptide nanocage or a polypeptide lattice with defined pore size. We designed, prepared and purified fusion protein between the N-terminal gyrase subunit B dimerization domain and trimerization domains composed of CutA1 and foldon. The main advantage of this system is that we can regulate the assembly and disassembly of this material since we can control the dimerization state of the gyrase B domain by adding aminocoumarin antibiotics novobiocin and coumermycin. Both of them bind to the same site on gyrase subunit B. Coumermycin is a pseudodimer and causes gyrase B dimerization while novobiocin binds competitively to the same binding site and disrupts coumermycin-induced dimers. In this way we can assemble or disassemble polypeptide nanostructures, which could be particularly useful to enmesh compounds such as biological drugs or any other application, where controlled release is desired. We demonstrated the ability of the material to assemble and disassemble in the presence of coumermycin and novobiocin respectively and observed the porous structure of the material at the nanoscale by transmission electron microscope.
