mRNA display selection of a high-affinity, Bcl-XL-specific binding peptide

Nobutaka Matsumura, Toru Tsuji, Takeshi Sumida, Masahito Kokubo, Michiko Onimaru, Nobuhide Doi, Hideaki Takashima, Etsuko Miyamoto-Sato, Hiroshi Yanagawa

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Bcl-XL, an antiapoptotic member of the Bcl-2 family, is a mitochondrial protein that inhibits activation of Bax and Bak, which commit the cell to apoptosis, and it therefore represents a potential target for drug discovery. Peptides have potential as therapeutic molecules because they can be designed to engage a larger portion of the target protein with higher specificity. In the present study, we selected 16-mer peptides that interact with Bcl-XL from random and degenerate peptide libraries using mRNA display. The selected peptides have sequence similarity with the Bcl-2 family BH3 domains, and one of them has higher affinity (IC50=0.9 μM) than Bak BH3 (IC50=11.8 μM) for Bcl-XL in vitro. We also found that GFP fusions of the selected peptides specifically interact with Bcl-XL, localize in mitochondria, and induce cell death. Further, a chimeric molecule, in which the BH3 domain of Bak protein was replaced with a selected peptide, retained the ability to bind specifically to Bcl-X L. These results demonstrate that this selected peptide specifically antagonizes the function of Bcl-XL and overcomes the effects of Bcl-XL in intact cells. We suggest that mRNA display is a powerful technique to identify peptide inhibitors with high affinity and specificity for disease-related proteins.

Original languageEnglish
Pages (from-to)2201-2210
Number of pages10
JournalFASEB Journal
Volume24
Issue number7
DOIs
Publication statusPublished - 2010 Jul

Fingerprint

Display devices
Messenger RNA
Peptides
Inhibitory Concentration 50
bcl-2 Homologous Antagonist-Killer Protein
Peptide Library
Mitochondria
Molecules
Mitochondrial Proteins
Cell death
Drug Discovery
Proteins
Cell Death
Fusion reactions
Chemical activation
Apoptosis
Therapeutics

Keywords

  • Bcl-2 family
  • BH3 domain
  • Therapeutic peptide

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology

Cite this

Matsumura, N., Tsuji, T., Sumida, T., Kokubo, M., Onimaru, M., Doi, N., ... Yanagawa, H. (2010). mRNA display selection of a high-affinity, Bcl-XL-specific binding peptide. FASEB Journal, 24(7), 2201-2210. https://doi.org/10.1096/fj.09-143008

mRNA display selection of a high-affinity, Bcl-XL-specific binding peptide. / Matsumura, Nobutaka; Tsuji, Toru; Sumida, Takeshi; Kokubo, Masahito; Onimaru, Michiko; Doi, Nobuhide; Takashima, Hideaki; Miyamoto-Sato, Etsuko; Yanagawa, Hiroshi.

In: FASEB Journal, Vol. 24, No. 7, 07.2010, p. 2201-2210.

Research output: Contribution to journalArticle

Matsumura, N, Tsuji, T, Sumida, T, Kokubo, M, Onimaru, M, Doi, N, Takashima, H, Miyamoto-Sato, E & Yanagawa, H 2010, 'mRNA display selection of a high-affinity, Bcl-XL-specific binding peptide', FASEB Journal, vol. 24, no. 7, pp. 2201-2210. https://doi.org/10.1096/fj.09-143008
Matsumura N, Tsuji T, Sumida T, Kokubo M, Onimaru M, Doi N et al. mRNA display selection of a high-affinity, Bcl-XL-specific binding peptide. FASEB Journal. 2010 Jul;24(7):2201-2210. https://doi.org/10.1096/fj.09-143008
Matsumura, Nobutaka ; Tsuji, Toru ; Sumida, Takeshi ; Kokubo, Masahito ; Onimaru, Michiko ; Doi, Nobuhide ; Takashima, Hideaki ; Miyamoto-Sato, Etsuko ; Yanagawa, Hiroshi. / mRNA display selection of a high-affinity, Bcl-XL-specific binding peptide. In: FASEB Journal. 2010 ; Vol. 24, No. 7. pp. 2201-2210.
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