Peptide Ligands for Pro-survival Protein Bfl-1 from Computationally Guided Library Screening
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 peptid...
| Main Authors: | , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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American Chemical Society (ACS)
2015
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| Online Access: | http://hdl.handle.net/1721.1/96522 https://orcid.org/0000-0003-4074-8980 |
| _version_ | 1826211591944667136 |
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| author | Dutta, Sanjib Chen, T. Scott Keating, Amy E. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Dutta, Sanjib Chen, T. Scott Keating, Amy E. |
| author_sort | Dutta, Sanjib |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T15:08:25Z |
| format | Article |
| id | mit-1721.1/96522 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:08:25Z |
| publishDate | 2015 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/965222022-09-29T12:56:14Z Peptide Ligands for Pro-survival Protein Bfl-1 from Computationally Guided Library Screening Dutta, Sanjib Chen, T. Scott Keating, Amy E. Massachusetts Institute of Technology. Department of Biology Dutta, Sanjib Chen, T. Scott Keating, Amy E. 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. National Institute of General Medical Sciences (U.S.) (Award GM084181) National Institute of General Medical Sciences (U.S.) (Award P50-GM68762) 2015-04-10T17:43:28Z 2015-04-10T17:43:28Z 2013-01 2012-12 Article http://purl.org/eprint/type/JournalArticle 1554-8929 1554-8937 http://hdl.handle.net/1721.1/96522 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. https://orcid.org/0000-0003-4074-8980 en_US http://dx.doi.org/10.1021/cb300679a ACS Chemical Biology Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society (ACS) PMC |
| spellingShingle | Dutta, Sanjib Chen, T. Scott Keating, Amy E. Peptide Ligands for Pro-survival Protein Bfl-1 from Computationally Guided Library Screening |
| title | Peptide Ligands for Pro-survival Protein Bfl-1 from Computationally Guided Library Screening |
| title_full | Peptide Ligands for Pro-survival Protein Bfl-1 from Computationally Guided Library Screening |
| title_fullStr | Peptide Ligands for Pro-survival Protein Bfl-1 from Computationally Guided Library Screening |
| title_full_unstemmed | Peptide Ligands for Pro-survival Protein Bfl-1 from Computationally Guided Library Screening |
| title_short | Peptide Ligands for Pro-survival Protein Bfl-1 from Computationally Guided Library Screening |
| title_sort | peptide ligands for pro survival protein bfl 1 from computationally guided library screening |
| url | http://hdl.handle.net/1721.1/96522 https://orcid.org/0000-0003-4074-8980 |
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