Structure and self-assembly of negatively supercharged protein cages

Eita Sasaki, Donald Hilvert

Research output: Contribution to journalReview articlepeer-review

1 Citation (Scopus)


Proteins are excellent materials for constructing nano-to micro-meter sized compartments. For example, in nature, hollow spherical shells made of proteins, called protein cages, are widespread. Prominent examples include viruses, fer-ritins, carboxysomes, and others. Protein cages designed and engineered in the laboratory have also gained recent attention because of their potential use in synthetic biology, materials science, and medicine. Here, we show that engineered variants of lumazine synthase (LS) from Aquifex aeolicus self-assemble into porous shell-like structures, with striking size-expansion from the original dodecahedron composed of 12 pentamer subunits. Cryo-electron microscopy (EM) analysis has revealed that pentamers are the basic assembly units, although small conformational changes in each sub-unit lead to final expanded architectures composed of 36 and 72 pentamers. The underlying conformational changes likely arise from electrostatic repulsion between anionic residues originally introduced at the lumenal surface of the LS cage to encapsulate positively charged guest molecules. The plastic nature of the LS cage structure was also explored using a positively supercharged variant of the green fluorescent protein GFP(+36) as an assembly mediator. By controlling the favorable electrostatic interactions between the negatively charged LS cage and the positively charged mediator, multishell structures were created, as previously observed in some virus-like particles. These results highlight the potential of engineered LS cages for various future applications, including drug delivery and bioimaging.

Original languageEnglish
Pages (from-to)199-208
Number of pages10
JournalYakugaku Zasshi
Issue number2
Publication statusPublished - 2019
Externally publishedYes


  • Cryo-electron microscopy
  • Lumazine synthase
  • Protein cage
  • Self-assembly
  • Supercharged protein
  • Virus-like particle

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science


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