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Nat Chem. 2019 Mar;11(3):204-212. doi: 10.1038/s41557-018-0196-3. Epub 2019 Jan 14.

Supercharging enables organized assembly of synthetic biomolecules.

Author information

1
Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, TX, USA.
2
Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX, USA.
3
Department of Molecular Biosciences, University of Texas at Austin, Austin, TX, USA.
4
Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, USA.
5
US Army Research Laboratory - South, Austin, TX, USA.
6
Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea.
7
Department of Materials Science & Engineering, University of Michigan, Ann Arbor, MI, USA.
8
Biointerfaces Institute, University of Michigan, Ann Arbor, MI, USA.
9
Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, TX, USA. dtaylor@utexas.edu.
10
Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX, USA. dtaylor@utexas.edu.
11
Department of Molecular Biosciences, University of Texas at Austin, Austin, TX, USA. dtaylor@utexas.edu.
12
Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, TX, USA. andy.ellington@mail.utexas.edu.
13
Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX, USA. andy.ellington@mail.utexas.edu.
14
Department of Molecular Biosciences, University of Texas at Austin, Austin, TX, USA. andy.ellington@mail.utexas.edu.

Abstract

Symmetrical protein oligomers are ubiquitous in biological systems and perform key structural and regulatory functions. However, there are few methods for constructing such oligomers. Here we have engineered completely synthetic, symmetrical oligomers by combining pairs of oppositely supercharged variants of a normally monomeric model protein through a strategy we term 'supercharged protein assembly' (SuPrA). We show that supercharged variants of green fluorescent protein can assemble into a variety of architectures including a well-defined symmetrical 16-mer structure that we solved using cryo-electron microscopy at 3.47 Å resolution. The 16-mer is composed of two stacked rings of octamers, in which the octamers contain supercharged proteins of alternating charges, and interactions within and between the rings are mediated by a variety of specific electrostatic contacts. The ready assembly of this structure suggests that combining oppositely supercharged pairs of protein variants may provide broad opportunities for generating novel architectures via otherwise unprogrammed interactions.

PMID:
30643229
DOI:
10.1038/s41557-018-0196-3
[Indexed for MEDLINE]

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