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Nat Biotechnol. 2017 Nov;35(11):1094-1101. doi: 10.1038/nbt.3994. Epub 2017 Oct 16.

Design of coiled-coil protein-origami cages that self-assemble in vitro and in vivo.

Author information

1
Department of Synthetic Biology and Immunology, National Institute of Chemistry, Ljubljana, Slovenia.
2
Graduate School of Biomedicine, University of Ljubljana, Ljubljana, Slovenia.
3
EN-FIST Centre of Excellence, Ljubljana, Slovenia.
4
Doctoral Study Programme in Chemical Sciences, University of Ljubljana, Ljubljana, Slovenia.
5
FAMINT, University of Primorska, Koper and Institute of Mathematics and Physics, Ljubljana, Slovenia.
6
Center of Excellence for Molecular Food Sciences, Faculty of Chemistry, University of Belgrade, Serbia.
7
The European Synchrotron Radiation Facility (ESRF), Grenoble, France.
8
School of Chemical and Physical Sciences, Keele Univerity, Keele, Staffordshire, UK.
9
Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain.

Abstract

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.

PMID:
29035374
DOI:
10.1038/nbt.3994
[Indexed for MEDLINE]

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