Truncated and helix-constrained peptides with high affinity and specificity for the cFos coiled-coil of AP-1.
Protein-based therapeutics feature large interacting surfaces. Protein folding endows structural stability to localised surface epitopes, imparting high affinity and target specificity upon interactions with binding partners. However, short synthetic peptides with sequences corresponding to such pro...
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| Language: | English |
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0059415&type=printable |
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| author | Tara Rao Gloria Ruiz-Gómez Timothy A Hill Huy N Hoang David P Fairlie Jody M Mason |
| author_facet | Tara Rao Gloria Ruiz-Gómez Timothy A Hill Huy N Hoang David P Fairlie Jody M Mason |
| author_sort | Tara Rao |
| collection | DOAJ |
| description | Protein-based therapeutics feature large interacting surfaces. Protein folding endows structural stability to localised surface epitopes, imparting high affinity and target specificity upon interactions with binding partners. However, short synthetic peptides with sequences corresponding to such protein epitopes are unstructured in water and promiscuously bind to proteins with low affinity and specificity. Here we combine structural stability and target specificity of proteins, with low cost and rapid synthesis of small molecules, towards meeting the significant challenge of binding coiled coil proteins in transcriptional regulation. By iteratively truncating a Jun-based peptide from 37 to 22 residues, strategically incorporating i→i+4 helix-inducing constraints, and positioning unnatural amino acids, we have produced short, water-stable, α-helical peptides that bind cFos. A three-dimensional NMR-derived structure for one peptide (24) confirmed a highly stable α-helix which was resistant to proteolytic degradation in serum. These short structured peptides are entropically pre-organized for binding with high affinity and specificity to cFos, a key component of the oncogenic transcriptional regulator Activator Protein-1 (AP-1). They competitively antagonized the cJun-cFos coiled-coil interaction. Truncating a Jun-based peptide from 37 to 22 residues decreased the binding enthalpy for cJun by ∼9 kcal/mol, but this was compensated by increased conformational entropy (TΔS ≤7.5 kcal/mol). This study demonstrates that rational design of short peptides constrained by α-helical cyclic pentapeptide modules is able to retain parental high helicity, as well as high affinity and specificity for cFos. These are important steps towards small antagonists of the cJun-cFos interaction that mediates gene transcription in cancer and inflammatory diseases. |
| format | Article |
| id | doaj-art-79d79d2f77d44a0fb8774105cc86847a |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-79d79d2f77d44a0fb8774105cc86847a2025-08-20T02:05:32ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0183e5941510.1371/journal.pone.0059415Truncated and helix-constrained peptides with high affinity and specificity for the cFos coiled-coil of AP-1.Tara RaoGloria Ruiz-GómezTimothy A HillHuy N HoangDavid P FairlieJody M MasonProtein-based therapeutics feature large interacting surfaces. Protein folding endows structural stability to localised surface epitopes, imparting high affinity and target specificity upon interactions with binding partners. However, short synthetic peptides with sequences corresponding to such protein epitopes are unstructured in water and promiscuously bind to proteins with low affinity and specificity. Here we combine structural stability and target specificity of proteins, with low cost and rapid synthesis of small molecules, towards meeting the significant challenge of binding coiled coil proteins in transcriptional regulation. By iteratively truncating a Jun-based peptide from 37 to 22 residues, strategically incorporating i→i+4 helix-inducing constraints, and positioning unnatural amino acids, we have produced short, water-stable, α-helical peptides that bind cFos. A three-dimensional NMR-derived structure for one peptide (24) confirmed a highly stable α-helix which was resistant to proteolytic degradation in serum. These short structured peptides are entropically pre-organized for binding with high affinity and specificity to cFos, a key component of the oncogenic transcriptional regulator Activator Protein-1 (AP-1). They competitively antagonized the cJun-cFos coiled-coil interaction. Truncating a Jun-based peptide from 37 to 22 residues decreased the binding enthalpy for cJun by ∼9 kcal/mol, but this was compensated by increased conformational entropy (TΔS ≤7.5 kcal/mol). This study demonstrates that rational design of short peptides constrained by α-helical cyclic pentapeptide modules is able to retain parental high helicity, as well as high affinity and specificity for cFos. These are important steps towards small antagonists of the cJun-cFos interaction that mediates gene transcription in cancer and inflammatory diseases.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0059415&type=printable |
| spellingShingle | Tara Rao Gloria Ruiz-Gómez Timothy A Hill Huy N Hoang David P Fairlie Jody M Mason Truncated and helix-constrained peptides with high affinity and specificity for the cFos coiled-coil of AP-1. PLoS ONE |
| title | Truncated and helix-constrained peptides with high affinity and specificity for the cFos coiled-coil of AP-1. |
| title_full | Truncated and helix-constrained peptides with high affinity and specificity for the cFos coiled-coil of AP-1. |
| title_fullStr | Truncated and helix-constrained peptides with high affinity and specificity for the cFos coiled-coil of AP-1. |
| title_full_unstemmed | Truncated and helix-constrained peptides with high affinity and specificity for the cFos coiled-coil of AP-1. |
| title_short | Truncated and helix-constrained peptides with high affinity and specificity for the cFos coiled-coil of AP-1. |
| title_sort | truncated and helix constrained peptides with high affinity and specificity for the cfos coiled coil of ap 1 |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0059415&type=printable |
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