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mRNA display selection of a high‐affinity, Bcl‐X L ‐specific binding peptide
Author(s) -
Matsumura Nobutaka,
Tsuji Toru,
Sumida Takeshi,
Kokubo Masahito,
Onimaru Michiko,
Doi Nobuhide,
Takashima Hideaki,
MiyamotoSato Etsuko,
Yanagawa Hiroshi
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.09-143008
Subject(s) - peptide , peptide library , biology , messenger rna , microbiology and biotechnology , function (biology) , apoptosis , peptide sequence , chemistry , biochemistry , gene
Bcl‐X L , 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‐X L 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 (IC 50 =0.9 µM) than Bak BH3 (IC 50 =11.8 µM) for Bcl‐X L in vitro. We also found that GFP fusions of the selected peptides specifically interact with Bcl‐X L , 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‐X L and overcomes the effects of Bcl‐X L in intact cells. We suggest that mRNA display is a powerful technique to identify peptide inhibitors with high affinity and specificity for diseaserelated proteins.—Matsumura, N., Tsuji, T., Sumida, T., Kokubo, M., Onimaru, M., Doi, N., Takashima, H., Miyamoto‐Sato, E., Yanagawa, H. mRNA display selection of a high‐affinity, Bcl‐X L ‐specific binding peptide. FASEBJ. 24, 2201–2210 (2010). www.fasebj.org