Self-Associating Peptides for Modular Bifunctional Conjugation of Tetramer Macromolecules in Living Cells
| Autor: | Agnès M. Cordonnier, Mariel Donzeau, Yves Nominé, Annick Dejaegere, Bruno Chatton, Frank Dietsch, Marc Vigneron, Guy Zuber, Laurent Bianchetti |
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| Přispěvatelé: | univOAK, Archive ouverte, Biotechnologie et signalisation cellulaire (BSC), Université de Strasbourg (UNISTRA)-Institut de recherche de l'Ecole de biotechnologie de Strasbourg (IREBS)-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2019 |
| Předmět: |
DNA Replication
Models Molecular heterotetramer Biomedical Engineering Pharmaceutical Science Bioengineering Sciences du Vivant [q-bio]/Biochimie Biologie Moléculaire Sciences du Vivant [q-bio]/Génétique chemistry.chemical_compound Protein Domains Tetramer Cell Line Tumor Proliferating Cell Nuclear Antigen Sciences du Vivant [q-bio]/Biologie cellulaire [SDV.BBM] Life Sciences [q-bio]/Biochemistry Molecular Biology Humans PCNA [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Bifunctional modularity Pharmacology business.industry Organic Chemistry food and beverages DNA non-covalent Modular design Combinatorial chemistry peptide bifunctional chemistry Covalent bond Protein Multimerization Tumor Suppressor Protein p53 Peptides business Biotechnology Macromolecule |
| Zdroj: | Bioconjugate Chemistry Bioconjugate Chemistry, 2019, 30 (6), pp.1734-1744. ⟨10.1021/acs.bioconjchem.9b00276⟩ |
| ISSN: | 1520-4812 1043-1802 |
| Popis: | Monitoring the assembly of macromolecules to design entities with novel properties can be achieved either chemically creating covalent bonds or by noncovalent connections using appropriate structural motifs. In this report, two self-associating peptides (named K3 and E3) that originate from p53 tetramerization domain were developed as tools for highly specific and noncovalent heterotetramerization of two biomolecules. The pairing/coupling preferences of K3 and E3 were first evaluated by molecular modeling data and confirmed using circular dichroism spectroscopy, size-exclusion chromatography, and biological assays. Regardless of the moieties fused to K3 and E3, these two peptides self-assembled into dimers of dimers to form bivalent heterotetrameric complexes that proved to be extremely stable inside living cells. The benefits of the multivalency in terms of avidity, specificity, and expanded functional activity were strikingly revealed when the proliferating cell nuclear antigen (PCNA), which is essential for DNA replication, was targeted using a heterotetramer presenting both an antibody fragment against PCNA and a specific PCNA binder peptide. In vitro heterotetramerization of these two known PCNA ligands increased their binding efficiencies to PCNA up to 80-fold compared to the best homotetramer counterpart. In cellulo, the heterotetramers were able to efficiently inhibit DNA replication and to trigger cell death. Altogether, we demonstrate that these two biselective self-assembling peptidic domains offer a versatile noncovalent conjugation method that can be easily implemented for protein engineering. papers2://publication/uuid/89537C97-09B4-4D8F-B955-FFAF0CF7E08A |
| Databáze: | OpenAIRE |
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