title: A Biochemical/Biophysical Assay Dyad for HTS-Compatible Triaging of Inhibitors of the HIV-1 Nef/Hck SH3 Interaction.
abstract: The current treatment regimens for HIV include over 20 anti-retrovirals. However, adverse drug effects and the emergence of drug resistance necessitates the continued improvement of the existing drug classes as well as the development of novel drugs that target as yet therapeutically unexploited viral and cellular pathways. Here we demonstrate a strategy for the discovery of protein-protein interaction inhibitors of the viral pathogenicity factor HIV-1 Nef and its interaction with the host factor SH3. A combination of a time-resolved fluorescence resonance energy resonance energy transfer-based assay and a label-free resonant waveguide grating-based assay was optimized for high-throughput screening formats.
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Breuer
authors.affiliation: Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Dr, San Diego, CA 92121, USA.
authors.forename: Sheryll
authors.initials: S
authors.lastname: Espinola
authors.affiliation: CRCM, CNRS UMR7258, Laboratory of Integrative Structural and Chemical Biology (ISCB); INSERM, U1068; Institut Paoli-Calmettes; Aix-Marseille Université, UM105, F-13009, Marseille, France.
authors.forename: Xavier
authors.initials: X
authors.lastname: Morelli
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Bruce E
authors.initials: BE
authors.lastname: Torbett
authors.affiliation: Department of Biochemistry and Molecular Biology, Unit 1000, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX 77030, USA ; King Abdullah University of Science and Technology (KAUST), Division of Biological and Environmental Sciencesand Engineering, Computational Bioscience Research Center, Thuwal 23955-6900, Saudi Arabia.
authors.forename: Stefan T
authors.initials: ST
authors.lastname: Arold
authors.affiliation: Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Dr, San Diego, CA 92121, USA.
authors.forename: Ingo H
authors.initials: IH
authors.lastname: Engels
keywords: HIV; HTS; Nef; SH3; TR-FRET.; label-free technology; protein-protein interaction inhibitor; resonant waveguide grating