title: Screening for Protein-Protein Interaction Inhibitors Using a Bioluminescence Resonance Energy Transfer (BRET)-Based Assay in Yeast.
abstract: The bioluminescence resonance energy transfer (BRET) technology is a widely used live cell-based method for monitoring protein-protein interactions as well as conformational changes within proteins or molecular complexes. Considering the emergence of protein-protein interactions as a new promising class of therapeutic targets, we have adapted the BRET method in budding yeast. In this technical note, we describe the advantages of using this simple eukaryotic model rather than mammalian cells to perform high-throughput screening of chemical compound collections: genetic tractability, tolerance to solvent, rapidity, and no need of expensive robotic systems. Here, the HDM2/p53 interaction, related to cancer, is used to highlight the interest of this technology in yeast. Sharing the protocol of this BRET-based assay with the scientific community will extend its application to other protein-protein interactions, even though it is toxic for mammalian cells, in order to discover promising therapeutic candidates.
authors.affiliation: 1 Sorbonne Universités, UPMC Univ Paris 06, CNRS USR3151, Protein Phosphorylation and Human Disease Unit, Station Biologique de Roscoff, Roscoff Cedex, France.|2 Université Bretagne Sud, FRE CNRS3744, IRDL, Vannes, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Corbel
authors.affiliation: 3 Laboratori di Biochimica e Biologia Molecolare, Dipartimento di Bioscienze, Università degli Studi di Parma, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Sartini
authors.affiliation: 3 Laboratori di Biochimica e Biologia Molecolare, Dipartimento di Bioscienze, Università degli Studi di Parma, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Levati
authors.affiliation: 1 Sorbonne Universités, UPMC Univ Paris 06, CNRS USR3151, Protein Phosphorylation and Human Disease Unit, Station Biologique de Roscoff, Roscoff Cedex, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Colas
authors.affiliation: 4 U892-INSERM Centre de Recherche en Cancérologie Nantes-Angers, Institut de Recherche en Santé de l'Université de Nantes, Nantes, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Maillet
authors.affiliation: 5 Université Lille, Inserm, Institut Pasteur de Lille, U1177-Drugs & Molecules for Living Systems, Lille, France.
authors.forename: Cyril
authors.initials: C
authors.lastname: Couturier
authors.affiliation: 3 Laboratori di Biochimica e Biologia Molecolare, Dipartimento di Bioscienze, Università degli Studi di Parma, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Montanini
authors.affiliation: 1 Sorbonne Universités, UPMC Univ Paris 06, CNRS USR3151, Protein Phosphorylation and Human Disease Unit, Station Biologique de Roscoff, Roscoff Cedex, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Bach
mesh_terms: D060807:Bioluminescence Resonance Energy Transfer Techniques; D055808:Drug Discovery / Q000379:methods*; D004353:Drug Evaluation, Preclinical; D005434:Flow Cytometry; D011485:Protein Binding / Q000187:drug effects*; D025941:Protein Interaction Mapping / Q000379:methods; D060066:Protein Interaction Maps; D057188:Workflow; D015003:Yeasts / Q000187:drug effects* / Q000378:metabolism*
keywords: HDM2/p53; bioluminescence resonance energy transfer (BRET); protein-protein interaction; yeast screening assays