RESEARCH ARTICLE


A Biochemical/Biophysical Assay Dyad for HTS-Compatible Triaging of Inhibitors of the HIV-1 Nef/Hck SH3 Interaction



Sebastian Breuer1, *, Sheryll Espinola2, Xavier Morelli3, Bruce E Torbett1, Stefan T Arold4, 5, Ingo H Engels2
1 Department of Molecular and Experimental Medicine, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA
2 Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Dr, San Diego, CA 92121, USA
3 CRCM, CNRS UMR7258, Laboratory of Integrative Structural and Chemical Biology (ISCB); INSERM, U1068; Institut Paoli‐Calmettes; Aix‐Marseille Université, UM105, F‐13009, Marseille, France
4 Department of Biochemistry and Molecular Biology, Unit 1000, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX 77030, USA
5 King Abdullah University of Science and Technology (KAUST), Division of Biological and Environmental Sciencesand Engineering, Computational Bioscience Research Center, Thuwal 23955-6900, Saudi Arabia


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© Breuer et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the Department of Molecular and Experimental Medicine, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA; Tel: +1 858 784 9976; Fax: + 1 858 784 7714; E-mail: breuer@scripps.edu


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.

Keywords: HIV, HTS, label-free technology, Nef, protein-protein interaction inhibitor, resonant waveguide grating, SH3, TR-FRET.