Design and Evolution of SnoopLigase for Unbreakable Biomolecular Connections

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Vaccination is one of the most effective ways to assure the health of humans and other animals and to ensure the security of the food supply. There is an urgent need to improve the way that vaccines are generated, because we lack effective vaccines to many of the most challenging diseases. Also, standard approaches to generate vaccines are slow compared to the speed that outbreaks of new diseases can occur. New vaccines are often developed from Virus-like particles (VLPs), which resemble viruses on the outside, but cannot cause disease because they do not have the virus genome inside. VLPs can be engineered to display a protein from a human or veterinary pathogen on their surface, to create an effective and safe vaccine. Decorating VLPs with such proteins from pathogens is a central problem for vaccine development. VLP decoration requires expensive trial-and-error experiments and has proved unfeasible with many protein targets. We have recently engineered a "bacterial superglue", termed SnoopLigase, which can catalyse the unbreakable linkage of one protein to another protein. This attachment is irreversible and applicable to a wide range of protein targets. SnoopLigase has proved useful in making enzymes more tolerant to harsh conditions and is likely to have diverse applications, including in assembly of diagnostic devices. In this proposal we will use evolution and computer-based design to engineer the next generation of SnoopLigase technology, so that it is faster, active in living cells (not just in the test-tube), and more potent at VLP decoration. SnoopLigase should be a general resource for scientists worldwide and speed up the creation of effective vaccines against major disease challenges.