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Protein Cages and Nanostructures Constructed from Protein Nanobuilding Blocks

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Protein Cages

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2671))

Abstract

Protein cages and nanostructures are promising biocompatible medical materials, such as vaccines and drug carriers. Recent advances in designed protein nanocages and nanostructures have opened up cutting-edge applications in the fields of synthetic biology and biopharmaceuticals. A simple approach for constructing self-assembling protein nanocages and nanostructures is the design of a fusion protein composed of two different proteins forming symmetric oligomers. In this chapter, we describe the design and methods of protein nanobuilding blocks (PN-Blocks) using a dimeric de novo protein WA20 to construct self-assembling protein cages and nanostructures. A protein nanobuilding block (PN-Block), WA20-foldon, was developed by fusing an intermolecularly folded dimeric de novo protein WA20 and a trimeric foldon domain from bacteriophage T4 fibritin. The WA20-foldon self-assembled into several oligomeric nanoarchitectures in multiples of 6-mer. De novo extender protein nanobuilding blocks (ePN-Blocks) were also developed by fusing tandemly two WA20 with various linkers, to construct self-assembling cyclized and extended chain-like nanostructures. These PN-Blocks would be useful for the construction of self-assembling protein cages and nanostructures and their potential applications in the future.

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Acknowledgments

We thank Prof. Michael Hecht at Princeton University for the kind gift of the expression plasmid of WA20. We thank Prof. Takaaki Sato, Mr. Kouichi Inano, and Dr. Keiichi Yanase at Shinshu University for help in SAXS experiments and analysis. We thank Prof. Nobutaka Shimizu and Photon Factory (PF) staff for help in synchrotron SAXS experiments which were performed at PF, KEK under the approval of PF program advisory committee (Proposal No. 2014G111, 2016G153, and 2016G606). We thank Prof. Nobuyasu Koga, Dr. Rie Koga, and Dr. Takahiro Kosugi at the Institute for Molecular Science (IMS) for help in SEC-MALS experiments. This work was supported by Joint Research of IMS (IMS program No. 603, 206, 221). This work was supported by JSPS Research Fellowships (DC2) and JSPS KAKENHI Grant Numbers JP14J10185 and JP16H06837 to N.K., and JSPS KAKENHI Grant Numbers JP24113707, JP24780097, JP16K05841, JP16H00761, JP17KK0104, and JP19H02522 to R.A.

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Authors and Affiliations

  1. Division of Materials Science, Graduate School of Science and Technology, Nara Institute of Science and Technology, Ikoma, Nara, Japan

    Naoya Kobayashi

  2. Department of Biomolecular Innovation, Institute for Biomedical Sciences, Shinshu University, Ueda, Nagano, Japan

    Ryoichi Arai

  3. Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Ueda, Nagano, Japan

    Ryoichi Arai

Authors
  1. Naoya Kobayashi
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  2. Ryoichi Arai
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Correspondence to Ryoichi Arai .

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Editors and Affiliations

  1. Tokyo Institute Of Technology, Yokohama, Japan

    Takafumi Ueno

  2. Chemical & Biomedical Enginering, Nanyang Technological University, Singapore, Singapore

    Sierin Lim

  3. Nanyang Technological University, Singapore, Singapore

    Kelin Xia

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Kobayashi, N., Arai, R. (2023). Protein Cages and Nanostructures Constructed from Protein Nanobuilding Blocks. In: Ueno, T., Lim, S., Xia, K. (eds) Protein Cages. Methods in Molecular Biology, vol 2671. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3222-2_4

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  • DOI: https://doi.org/10.1007/978-1-0716-3222-2_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3221-5

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