Bifunctional Nanotube Scaffolds for Diverse Ligands Are Purified Simply fromEscherichia coliStrains Coexpressing Two Functionalized Flagellar Genes

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Abstract
We functionalized Escherichia coli FliC flagellin proteins to form tailored nanotubes binding single types or pairs of ligands, including divalent cations, fluorescent antibodies, or biotin-avidin-linked moieties such as ferritins. The ratio of each tag in bifunctionalized flagella could be toggled extending their sophistication as nanoscaffolds. Tobacco Etch Virus (TEV) protease site-containing FliCs were cleaved by the cognate protease without filament disintegration, potentiating their use as removable nanolithography masks to deposit attached ligands by protease cleavage.