Hyperstable de novo protein with a dimeric bisecting topology

Naoya Kimura, Kenji Mochizuki, Koji Umezawa, Michael H. Hecht, Ryoichi Arai

Research output: Contribution to journalArticlepeer-review

10 Scopus citations


Recently, we designed and assembled protein nanobuilding blocks (PN-Blocks) from an intermolecularly folded dimeric de novo protein called WA20. Using this dimeric 4-helix bundle, we constructed a series of self-assembling supramolecular nanostructures including polyhedra and chain-type complexes. Here we describe the stabilization of WA20 by designing mutations that stabilize the helices and hydrophobic core. The redesigned proteins denature with substantially higher midpoints, with the most stable variant, called Super WA20 (SUWA), displaying an extremely high midpoint (Tm = 122 °C), much higher than the Tm of WA20 (75 °C). The crystal structure of SUWA reveals an intermolecularly folded dimer with bisecting U topology, similar to the parental WA20 structure, with two long α-helices of a protomer intertwined with the helices of another protomer. Molecular dynamics simulations demonstrate that the redesigned hydrophobic core in the center of SUWA significantly suppresses the deformation of helices observed in the same region of WA20, suggesting this is a critical factor stabilizing the SUWA structure. This hyperstable de novo protein is expected to be useful as nanoscale pillars of PN-Block components in new types of self-assembling nanoarchitectures.

Original languageEnglish (US)
Pages (from-to)254-259
Number of pages6
JournalACS Synthetic Biology
Issue number2
StatePublished - Feb 21 2020

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Biomedical Engineering


  • 4-helix bundle
  • Binary pattern
  • Bisecting U topology
  • De novo protein
  • Protein nanobuilding block
  • Protein stabilization


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