Plug-and-Play Streptavidin Hubs for Megaprotein Assembly and Nanoscale Control of Cell-Surface Triggering

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There is strength in numbers, even at the molecular level. Biological molecules in clusters often have effects that cannot be achieved by those molecules in isolation. It is well known how to cluster one biological molecule into teams of 4, using a protein called streptavidin. Streptavidin is one of the most widely used tools in biological research and biotechnology because of this efficient and stable clustering. In this proposal we will establish how to engineer streptavidin to build a range of larger clusters containing precise numbers of molecules, up to 33. As well as determining the rules for how to build unbreakable clusters of different size and orientation, our new clusters will be powerful tools for studying and controlling many biological events. Signals are often communicated to cells through clustering of proteins at their surface. Killer T cells patrol the body, scrutinising the cells they pass to determine whether that cell is infected or healthy. According to the organisation of the clustering of receptors on the T cell, the T cell may kill the target cell or leave it unharmed. We will apply our different clusters to understand how the T cell makes this crucial decision.