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MultiBacMam Bimolecular Fluorescence Complementation (BiFC) tool-kit identifies new small-molecule inhibitors of the CDK5-p25 protein-protein interaction (PPI)

Protein-protein interactions (PPIs) are at the core of virtually all biological processes in cells. Consequently, targeting PPIs is emerging at the forefront of drug discovery. Cellular assays which closely recapitulate native conditions in vivo are instrumental to understand how small molecule drug...

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Autores principales: Bellón-Echeverría, Itxaso, Carralot, Jean-Philippe, Del Rosario, Andrea Araujo, Kueng, Stephanie, Mauser, Harald, Schmid, Georg, Thoma, Ralf, Berger, Imre
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5865166/
https://www.ncbi.nlm.nih.gov/pubmed/29572554
http://dx.doi.org/10.1038/s41598-018-23516-x
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author Bellón-Echeverría, Itxaso
Carralot, Jean-Philippe
Del Rosario, Andrea Araujo
Kueng, Stephanie
Mauser, Harald
Schmid, Georg
Thoma, Ralf
Berger, Imre
author_facet Bellón-Echeverría, Itxaso
Carralot, Jean-Philippe
Del Rosario, Andrea Araujo
Kueng, Stephanie
Mauser, Harald
Schmid, Georg
Thoma, Ralf
Berger, Imre
author_sort Bellón-Echeverría, Itxaso
collection PubMed
description Protein-protein interactions (PPIs) are at the core of virtually all biological processes in cells. Consequently, targeting PPIs is emerging at the forefront of drug discovery. Cellular assays which closely recapitulate native conditions in vivo are instrumental to understand how small molecule drugs can modulate such interactions. We have integrated MultiBacMam, a baculovirus-based mammalian gene delivery tool we developed, with bimolecular fluorescence complementation (BiFC), giving rise to a highly efficient system for assay development, identification and characterization of PPI modulators. We used our system to analyze compounds impacting on CDK5-p25 PPI, which is implicated in numerous diseases including Alzheimer’s. We evaluated our tool-kit with the known inhibitor p5T, and we established a mini-screen to identify compounds that modulate this PPI in dose-response experiments. Finally, we discovered several compounds disrupting CDK5-p25 PPI, which had not been identified by other screening or structure-based methods before.
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spelling pubmed-58651662018-03-27 MultiBacMam Bimolecular Fluorescence Complementation (BiFC) tool-kit identifies new small-molecule inhibitors of the CDK5-p25 protein-protein interaction (PPI) Bellón-Echeverría, Itxaso Carralot, Jean-Philippe Del Rosario, Andrea Araujo Kueng, Stephanie Mauser, Harald Schmid, Georg Thoma, Ralf Berger, Imre Sci Rep Article Protein-protein interactions (PPIs) are at the core of virtually all biological processes in cells. Consequently, targeting PPIs is emerging at the forefront of drug discovery. Cellular assays which closely recapitulate native conditions in vivo are instrumental to understand how small molecule drugs can modulate such interactions. We have integrated MultiBacMam, a baculovirus-based mammalian gene delivery tool we developed, with bimolecular fluorescence complementation (BiFC), giving rise to a highly efficient system for assay development, identification and characterization of PPI modulators. We used our system to analyze compounds impacting on CDK5-p25 PPI, which is implicated in numerous diseases including Alzheimer’s. We evaluated our tool-kit with the known inhibitor p5T, and we established a mini-screen to identify compounds that modulate this PPI in dose-response experiments. Finally, we discovered several compounds disrupting CDK5-p25 PPI, which had not been identified by other screening or structure-based methods before. Nature Publishing Group UK 2018-03-23 /pmc/articles/PMC5865166/ /pubmed/29572554 http://dx.doi.org/10.1038/s41598-018-23516-x Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Bellón-Echeverría, Itxaso
Carralot, Jean-Philippe
Del Rosario, Andrea Araujo
Kueng, Stephanie
Mauser, Harald
Schmid, Georg
Thoma, Ralf
Berger, Imre
MultiBacMam Bimolecular Fluorescence Complementation (BiFC) tool-kit identifies new small-molecule inhibitors of the CDK5-p25 protein-protein interaction (PPI)
title MultiBacMam Bimolecular Fluorescence Complementation (BiFC) tool-kit identifies new small-molecule inhibitors of the CDK5-p25 protein-protein interaction (PPI)
title_full MultiBacMam Bimolecular Fluorescence Complementation (BiFC) tool-kit identifies new small-molecule inhibitors of the CDK5-p25 protein-protein interaction (PPI)
title_fullStr MultiBacMam Bimolecular Fluorescence Complementation (BiFC) tool-kit identifies new small-molecule inhibitors of the CDK5-p25 protein-protein interaction (PPI)
title_full_unstemmed MultiBacMam Bimolecular Fluorescence Complementation (BiFC) tool-kit identifies new small-molecule inhibitors of the CDK5-p25 protein-protein interaction (PPI)
title_short MultiBacMam Bimolecular Fluorescence Complementation (BiFC) tool-kit identifies new small-molecule inhibitors of the CDK5-p25 protein-protein interaction (PPI)
title_sort multibacmam bimolecular fluorescence complementation (bifc) tool-kit identifies new small-molecule inhibitors of the cdk5-p25 protein-protein interaction (ppi)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5865166/
https://www.ncbi.nlm.nih.gov/pubmed/29572554
http://dx.doi.org/10.1038/s41598-018-23516-x
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