Type III secretion filaments as templates for metallic nanostructure synthesis

Nanostructured materials can be interfaced with living cells to enable unique chemical and biological outcomes. However, it is challenging to precisely control the shape and chemical composition of submillimeter sized, cell-associated materials. In this protocol, we describe how to genetically modify and isolate a self-assembling filament protein from Salmonella enterica, PrgI, to bind Au nanoparticles. Au-conjugated filaments can be chemically reduced in vitro to form contiguous wires and networks that are several micrometers in length. We also describe a strategy to assemble PrgI-based filaments on live cells, which can then be sheared or remain tethered to cells for gold conjugation. These methods form the basis of a strategy for interactions between inorganic and organic systems, and could be expanded to introduce interactions with other metal nanoparticles for which peptide binding partners are known.