Chemical Proteomics for Target Discovery of Head-to-Tail Cyclized Mini-Proteins
Target deconvolution is one of the most challenging tasks in drug discovery, but a key step in drug development. In contrast to small molecules, there is a lack of validated and robust methodologies for target elucidation of peptides. In particular, it is difficult to apply these methods to cyclic a...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2017-10-01
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| Series: | Frontiers in Chemistry |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fchem.2017.00073/full |
| _version_ | 1811211547298496512 |
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| author | Roland Hellinger Kathrin Thell Mina Vasileva Taj Muhammad Sunithi Gunasekera Daniel Kümmel Ulf Göransson Christian W. Becker Christian W. Gruber Christian W. Gruber |
| author_facet | Roland Hellinger Kathrin Thell Mina Vasileva Taj Muhammad Sunithi Gunasekera Daniel Kümmel Ulf Göransson Christian W. Becker Christian W. Gruber Christian W. Gruber |
| author_sort | Roland Hellinger |
| collection | DOAJ |
| description | Target deconvolution is one of the most challenging tasks in drug discovery, but a key step in drug development. In contrast to small molecules, there is a lack of validated and robust methodologies for target elucidation of peptides. In particular, it is difficult to apply these methods to cyclic and cysteine-stabilized peptides since they exhibit reduced amenability to chemical modification and affinity capture; however, such ribosomally synthesized and post-translationally modified peptide natural products are rich sources of promising drug candidates. For example, plant-derived circular peptides called cyclotides have recently attracted much attention due to their immunosuppressive effects and oral activity in the treatment of multiple sclerosis in mice, but their molecular target has hitherto not been reported. In this study, a chemical proteomics approach using photo-affinity crosslinking was developed to determine a target for the circular peptide [T20K]kalata B1. Using this prototypic nature-derived peptide enabled the identification of a possible functional modulation of 14-3-3 proteins. This biochemical interaction was validated via competition pull down assays as well as a cellular reporter assay indicating an effect on 14-3-3-dependent transcriptional activity. As proof of concept, the presented approach may be applicable for target elucidation of various cyclic peptides and mini-proteins, in particular cyclotides, which represent a promising class of molecules in drug discovery and development. |
| first_indexed | 2024-04-12T05:15:52Z |
| format | Article |
| id | doaj.art-57d35bad86d74a89b4e3a9d1ca88333b |
| institution | Directory Open Access Journal |
| issn | 2296-2646 |
| language | English |
| last_indexed | 2024-04-12T05:15:52Z |
| publishDate | 2017-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Chemistry |
| spelling | doaj.art-57d35bad86d74a89b4e3a9d1ca88333b2022-12-22T03:46:38ZengFrontiers Media S.A.Frontiers in Chemistry2296-26462017-10-01510.3389/fchem.2017.00073296256Chemical Proteomics for Target Discovery of Head-to-Tail Cyclized Mini-ProteinsRoland Hellinger0Kathrin Thell1Mina Vasileva2Taj Muhammad3Sunithi Gunasekera4Daniel Kümmel5Ulf Göransson6Christian W. Becker7Christian W. Gruber8Christian W. Gruber9Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, AustriaCenter for Physiology and Pharmacology, Medical University of Vienna, Vienna, AustriaCenter for Physiology and Pharmacology, Medical University of Vienna, Vienna, AustriaDivision of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Uppsala, SwedenDivision of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Uppsala, SwedenSchool of Biology/Chemistry, University of Osnabrück, Osnabrück, GermanyDivision of Pharmacognosy, Department of Medicinal Chemistry, Uppsala University, Uppsala, SwedenDepartment of Chemistry, Institute of Biological Chemistry, University of Vienna, Vienna, AustriaCenter for Physiology and Pharmacology, Medical University of Vienna, Vienna, AustriaSchool of Biomedical Sciences, Faculty of Medicine, University of Queensland, St. Lucia, QLD, AustraliaTarget deconvolution is one of the most challenging tasks in drug discovery, but a key step in drug development. In contrast to small molecules, there is a lack of validated and robust methodologies for target elucidation of peptides. In particular, it is difficult to apply these methods to cyclic and cysteine-stabilized peptides since they exhibit reduced amenability to chemical modification and affinity capture; however, such ribosomally synthesized and post-translationally modified peptide natural products are rich sources of promising drug candidates. For example, plant-derived circular peptides called cyclotides have recently attracted much attention due to their immunosuppressive effects and oral activity in the treatment of multiple sclerosis in mice, but their molecular target has hitherto not been reported. In this study, a chemical proteomics approach using photo-affinity crosslinking was developed to determine a target for the circular peptide [T20K]kalata B1. Using this prototypic nature-derived peptide enabled the identification of a possible functional modulation of 14-3-3 proteins. This biochemical interaction was validated via competition pull down assays as well as a cellular reporter assay indicating an effect on 14-3-3-dependent transcriptional activity. As proof of concept, the presented approach may be applicable for target elucidation of various cyclic peptides and mini-proteins, in particular cyclotides, which represent a promising class of molecules in drug discovery and development.http://journal.frontiersin.org/article/10.3389/fchem.2017.00073/fullcyclotidescyclic proteinchemical proteomicspeptide-protein interactionphoto-affinity labeling |
| spellingShingle | Roland Hellinger Kathrin Thell Mina Vasileva Taj Muhammad Sunithi Gunasekera Daniel Kümmel Ulf Göransson Christian W. Becker Christian W. Gruber Christian W. Gruber Chemical Proteomics for Target Discovery of Head-to-Tail Cyclized Mini-Proteins Frontiers in Chemistry cyclotides cyclic protein chemical proteomics peptide-protein interaction photo-affinity labeling |
| title | Chemical Proteomics for Target Discovery of Head-to-Tail Cyclized Mini-Proteins |
| title_full | Chemical Proteomics for Target Discovery of Head-to-Tail Cyclized Mini-Proteins |
| title_fullStr | Chemical Proteomics for Target Discovery of Head-to-Tail Cyclized Mini-Proteins |
| title_full_unstemmed | Chemical Proteomics for Target Discovery of Head-to-Tail Cyclized Mini-Proteins |
| title_short | Chemical Proteomics for Target Discovery of Head-to-Tail Cyclized Mini-Proteins |
| title_sort | chemical proteomics for target discovery of head to tail cyclized mini proteins |
| topic | cyclotides cyclic protein chemical proteomics peptide-protein interaction photo-affinity labeling |
| url | http://journal.frontiersin.org/article/10.3389/fchem.2017.00073/full |
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