Mimicking the evolution of a thermally stable monomeric four-helix bundle by fusion of four identical single-helix peptides

Satoshi Akanuma, Taku Matsuba, Emi Ueno, Naoki Umeda, Akihiko Yamagishi

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Internal symmetry is a common feature of the tertiary structures of proteins and protein domains. Probably, because the genes of homo-oligomeric proteins duplicated and fused, their evolutionary descendants are proteins with internal symmetry. To identify any advantages that cause monomeric proteins with internal symmetry to be selected evolutionarily, we characterized some of the physical properties of a recombinant protein with a sequence consisting of two tandemly fused copies of the Escherichia coli Lac repressor C-terminal α-helix. This polypeptide exists in solution mainly as dimer that likely maintains a four-helix bundle motif. Thermal unfolding experiments demonstrate that the protein is considerably more stable at elevated temperatures than is a homotetramer consisting of four non-covalently associated copies of a 21-residue polypeptide similar in sequence to that of the Lac repressor C-terminal α-helix. A tandem duplication of our helix-loop-helix polypeptide yields an even more thermally stable protein. Our results exemplify the concept that fusion of non-covalently assembled polypeptide chains leads to enhanced protein stability. Herein, we discuss how our work relates to the evolutionary selective-advantages realized when symmetrical homo-oligomers evolve into monomers. Moreover, our thermally stable single-chain four-helix bundle protein may provide a robust scaffold for development of new biomaterials.

Original languageEnglish
Pages (from-to)371-379
Number of pages9
JournalJournal of Biochemistry
Volume147
Issue number3
DOIs
Publication statusPublished - 2010 Mar
Externally publishedYes

Fingerprint

Fusion reactions
Peptides
Lac Repressors
Proteins
Protein Stability
Biocompatible Materials
Tertiary Protein Structure
Recombinant Proteins
Hot Temperature
Escherichia coli
Oligomers
Scaffolds
Dimers
Temperature
Physical properties
Genes
Monomers
Experiments

Keywords

  • Four-helix bundle
  • Fusion
  • Internal symmetry
  • Tandem duplication
  • Thermal stability

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Mimicking the evolution of a thermally stable monomeric four-helix bundle by fusion of four identical single-helix peptides. / Akanuma, Satoshi; Matsuba, Taku; Ueno, Emi; Umeda, Naoki; Yamagishi, Akihiko.

In: Journal of Biochemistry, Vol. 147, No. 3, 03.2010, p. 371-379.

Research output: Contribution to journalArticle

Akanuma, Satoshi ; Matsuba, Taku ; Ueno, Emi ; Umeda, Naoki ; Yamagishi, Akihiko. / Mimicking the evolution of a thermally stable monomeric four-helix bundle by fusion of four identical single-helix peptides. In: Journal of Biochemistry. 2010 ; Vol. 147, No. 3. pp. 371-379.
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