title: Rapid screening of protein-protein interaction inhibitors using the protease exclusion assay.
abstract: We have previously developed a sensitive and modular homogenous biosensor system using peptides to detect target ligands. By transposing the basic mechanistic principle of the nuclease protection assay into this biosensor framework, we have developed the protease exclusion (PE) assay which can discern antagonists of protein-protein interactions in a rapid, single-step format. We demonstrate the concept with multiple protein-peptide pairs and validate the method by successfully screening a small molecule library for compounds capable of inhibiting the therapeutically relevant p53-Mdm2 interaction. The Protease Exclusion method adds to the compendium of assays available for rapid analyte detection and is particularly suited for drug screening applications.
authors: p53Lab (A*STAR), 8A Biomedical Grove, #06-04/05 Neuros, Singapore 138648, Singapore. Electronic address: SNirantar@p53Lab.a-star.edu.sg.
authors: Saurabh R
authors: SR
authors: Nirantar
authors: p53Lab (A*STAR), 8A Biomedical Grove, #06-04/05 Neuros, Singapore 138648, Singapore.
authors: Xiang
authors: X
authors: Li
authors: p53Lab (A*STAR), 8A Biomedical Grove, #06-04/05 Neuros, Singapore 138648, Singapore.
authors: Jia Wei
authors: JW
authors: Siau
authors: p53Lab (A*STAR), 8A Biomedical Grove, #06-04/05 Neuros, Singapore 138648, Singapore. Electronic address: faridg@p53Lab.a-star.edu.sg.
authors: Farid J
authors: FJ
authors: Ghadessy
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005453:Fluorescence; D057166:High-Throughput Screening Assays / Q000379:methods; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps / Q000187:drug effects*; D051736:Proto-Oncogene Proteins c-mdm2 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D010455:Peptides; D054852:Small Molecule Libraries; D016159:Tumor Suppressor Protein p53; D051736:Proto-Oncogene Proteins c-mdm2
keywords: Biosensor; High throughput screening; P53–Mdm2 interaction; Protein–protein interaction