In-solution enrichment identifies peptide inhibitors of protein–protein interactions
The use of competitive inhibitors to disrupt protein–protein interactions (PPIs) holds great promise for the treatment of disease. However, the discovery of high-affinity inhibitors can be a challenge. Here we report a platform for improving the affinity of peptide-based PPI inhibitors using non-can...
| Main Authors: | , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
Springer Nature
2020
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| Online Access: | https://hdl.handle.net/1721.1/123681 |
| _version_ | 1826207906210512896 |
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| author | Touti, Faycal Gates, Zachary P Bandyopadhyay, Anupam Lautrette, Guillaume Pentelute, Bradley L. |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Touti, Faycal Gates, Zachary P Bandyopadhyay, Anupam Lautrette, Guillaume Pentelute, Bradley L. |
| author_sort | Touti, Faycal |
| collection | MIT |
| description | The use of competitive inhibitors to disrupt protein–protein interactions (PPIs) holds great promise for the treatment of disease. However, the discovery of high-affinity inhibitors can be a challenge. Here we report a platform for improving the affinity of peptide-based PPI inhibitors using non-canonical amino acids. The platform utilizes size exclusion-based enrichment from pools of synthetic peptides (1.5–4 kDa) and liquid chromatography-tandem mass spectrometry-based peptide sequencing to identify high-affinity binders to protein targets, without the need for ‘reporter’ or ‘encoding’ tags. Using this approach—which is inherently selective for high-affinity binders—we realized gains in affinity of up to ~100- or ~30-fold for binders to the oncogenic ubiquitin ligase MDM2 or HIV capsid protein C-terminal domain, which inhibit MDM2–p53 interaction or HIV capsid protein C-terminal domain dimerization, respectively. Subsequent macrocyclization of select MDM2 inhibitors rendered them cell permeable and cytotoxic toward cancer cells, demonstrating the utility of the identified compounds as functional PPI inhibitors. Keywords: chemical libraries; chemical tools; peptides; screening |
| first_indexed | 2024-09-23T13:56:29Z |
| format | Article |
| id | mit-1721.1/123681 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T13:56:29Z |
| publishDate | 2020 |
| publisher | Springer Nature |
| record_format | dspace |
| spelling | mit-1721.1/1236812022-09-28T17:17:23Z In-solution enrichment identifies peptide inhibitors of protein–protein interactions Touti, Faycal Gates, Zachary P Bandyopadhyay, Anupam Lautrette, Guillaume Pentelute, Bradley L. Massachusetts Institute of Technology. Department of Chemistry Koch Institute for Integrative Cancer Research at MIT The use of competitive inhibitors to disrupt protein–protein interactions (PPIs) holds great promise for the treatment of disease. However, the discovery of high-affinity inhibitors can be a challenge. Here we report a platform for improving the affinity of peptide-based PPI inhibitors using non-canonical amino acids. The platform utilizes size exclusion-based enrichment from pools of synthetic peptides (1.5–4 kDa) and liquid chromatography-tandem mass spectrometry-based peptide sequencing to identify high-affinity binders to protein targets, without the need for ‘reporter’ or ‘encoding’ tags. Using this approach—which is inherently selective for high-affinity binders—we realized gains in affinity of up to ~100- or ~30-fold for binders to the oncogenic ubiquitin ligase MDM2 or HIV capsid protein C-terminal domain, which inhibit MDM2–p53 interaction or HIV capsid protein C-terminal domain dimerization, respectively. Subsequent macrocyclization of select MDM2 inhibitors rendered them cell permeable and cytotoxic toward cancer cells, demonstrating the utility of the identified compounds as functional PPI inhibitors. Keywords: chemical libraries; chemical tools; peptides; screening Bettencourt Schueller Foundation (Award (LT000745/2014-C) United States. Defense Advanced Research Projects Agency (Award 023504-001) National Institute of General Medical Sciences (U.S.) (Grant 5-R01-GM110535) Laboratoires Servier (Gidy, France) (Award 023504-001) 2020-01-24T16:51:48Z 2020-01-24T16:51:48Z 2019-03-18 2017-11-02 2020-01-02T18:18:52Z Article http://purl.org/eprint/type/JournalArticle 1552-4450 1552-4469 https://hdl.handle.net/1721.1/123681 Touti, Fayçal et al. "In-solution enrichment identifies peptide inhibitors of protein–protein interactions." Nature Chemical Biology 15, 4 (April 2019): 410–418 © 2019 The Author(s) en http://dx.doi.org/10.1038/s41589-019-0245-2 Nature 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 Springer Nature PMC |
| spellingShingle | Touti, Faycal Gates, Zachary P Bandyopadhyay, Anupam Lautrette, Guillaume Pentelute, Bradley L. In-solution enrichment identifies peptide inhibitors of protein–protein interactions |
| title | In-solution enrichment identifies peptide inhibitors of protein–protein interactions |
| title_full | In-solution enrichment identifies peptide inhibitors of protein–protein interactions |
| title_fullStr | In-solution enrichment identifies peptide inhibitors of protein–protein interactions |
| title_full_unstemmed | In-solution enrichment identifies peptide inhibitors of protein–protein interactions |
| title_short | In-solution enrichment identifies peptide inhibitors of protein–protein interactions |
| title_sort | in solution enrichment identifies peptide inhibitors of protein protein interactions |
| url | https://hdl.handle.net/1721.1/123681 |
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