Rational engineering of self-assembling thermo-sensitive protein complexes and their applications

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This industrial biotechnology project relies on the biological resources and materials such as SNARE proteins-inspired molecular self-assembling system for producing thermosensitive pharmaceutical precursors, drug carriers and theranostic nanoparticles. Naturally occurring SNARE proteins drive fusion of cellular membranes in every eukaryotic cell. Using these we have developed a new molecular stapling method which allows site-oriented, non-covalent self-assembly of recombinant proteins (WO2011/018665A1). Such rationally engineered supramolecular self-assembling system is superior to all protein self-assembling systems known due to its resistance to chaotropic agents, strong detergents, proteases, elevated temperatures as well as its cell-penetrating properties. However the use of SNARES for drug delivery is hindered by their ability to interfere with native SNARES and the process of exocytosis. Our research effort is aimed at resolving this remaining problem. The overall aim of this research is engineer an expanded range of SNARE-inspired proteins which are capable of controlled self-assembly and dis-assembly and which do not interfere with native SNARES and are therefore suitable for in vivo applications. Two projects are available. One objective of this project is to identify key features of the expanded range of engineered SNARE-inspired proteins, responsible for the superior structural stability of SNAREs. Our methodology includes molecular modelling, computation analysis, recombinant protein expression, biochemical, biophysical, structural and thermal analyses. We will also test the effect of molecular cargo on the rates of self-assembly and the overall stability of these protein complexes. This project is a collaborative research between Mikhail Soloviev (Royal Holloway University of London), Enrico Ferrari (University of Lincoln) and Giuliano Siligardi (Diamond Light Source, Harwell Science Campus). Research will be conducted at all three sites. Another objective of this project is to engineer protein-based nanoparticles for targeted drug delivery. Our methodology includes molecular modelling, computation analysis, recombinant protein expression, biochemical and thermal analyses. We will also test these nanoparticles for their compatibility with a range of lipids and their effect on the thermal stability properties of our combinatorial bioparticle system. This project is a collaborative research between Mikhail Soloviev (Royal Holloway University of London) and Xiao Yun Xu (Imperial College London).