Peptide Ligands for Pro-survival Protein Bfl-1 from Computationally Guided Library Screening
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Keating_Peptide ligands.pdf
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Author(s)
Dutta, Sanjib
Chen, T. Scott
Keating, Amy E.
Date Issued
January 2013
Journal
ACS Chemical Biology
Publisher
American Chemical Society (ACS)
Citation
Dutta, Sanjib, T. Scott Chen, and Amy E. Keating. “Peptide Ligands for Pro-Survival Protein Bfl-1 from Computationally Guided Library Screening.” ACS Chemical Biology 8, no. 4 (April 19, 2013): 778–788.
Version
Author's final manuscript
Abstract
Pro-survival members of the Bcl-2 protein family inhibit cell death by binding short helical BH3 motifs in pro-apoptotic proteins. Mammalian pro-survival proteins Bcl-x[subscript L], Bcl-2, Bcl-w, Mcl-1, and Bfl-1 bind with varying affinities and specificities to native BH3 motifs, engineered peptides, and small molecules. Biophysical studies have determined interaction patterns for these proteins, particularly for the most-studied family members Bcl-x[subscript L] and Mcl-1. Bfl-1 is a pro-survival protein implicated in preventing apoptosis in leukemia, lymphoma, and melanoma. Although Bfl-1 is a promising therapeutic target, relatively little is known about its binding preferences. We explored the binding of Bfl-1 to BH3-like peptides by screening a peptide library that was designed to sample a high degree of relevant sequence diversity. Screening using yeast-surface display led to several novel high-affinity Bfl-1 binders and to thousands of putative binders identified through deep sequencing. Further screening for specificity led to identification of a peptide that bound to Bfl-1 with K[subscript d] < 1 nM and very slow dissociation from Bfl-1 compared to other pro-survival Bcl-2 family members. A point mutation in this sequence gave a peptide with ~50 nM affinity for Bfl-1 that was selective for Bfl-1 in equilibrium binding assays. Analysis of engineered Bfl-1 binders deepens our understanding of how the binding profiles of pro-survival proteins differ and may guide the development of targeted Bfl-1 inhibitors.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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DOI of Published Version
https://doi.org/10.1021/cb300679a
