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Design of coiled-coil protein-origami cages that self-assemble in vitro and in vivo
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Design of coiled-coil protein-origami cages that self-assemble in vitro and in vivo
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Design of coiled-coil protein-origami cages that self-assemble in vitro and in vivo
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Journal Article
Design of coiled-coil protein-origami cages that self-assemble in vitro and in vivo
2017
Overview
Protein origami using coiled-coil building blocks produces self-assembling polyhedral cages for diverse applications.
Polypeptides and polynucleotides are natural programmable biopolymers that can self-assemble into complex tertiary structures. We describe a system analogous to designed DNA nanostructures in which protein coiled-coil (CC) dimers serve as building blocks for modular
de novo
design of polyhedral protein cages that efficiently self-assemble
in vitro
and
in vivo
. We produced and characterized >20 single-chain protein cages in three shapes—tetrahedron, four-sided pyramid, and triangular prism—with the largest containing >700 amino-acid residues and measuring 11 nm in diameter. Their stability and folding kinetics were similar to those of natural proteins. Solution small-angle X-ray scattering (SAXS), electron microscopy (EM), and biophysical analysis confirmed agreement of the expressed structures with the designs. We also demonstrated self-assembly of a tetrahedral structure in bacteria, mammalian cells, and mice without evidence of inflammation. A semi-automated computational design platform and a toolbox of CC building modules are provided to enable the design of protein cages in any polyhedral shape.
