Using AlphaScreen(®) to Identify Small-Molecule Inhibitors Targeting a Conserved Host-Pathogen Interaction.
AlphaScreen(®) is a technology particularly suitable for bi-molecular inhibitor screening assays, e.g. using protein-protein interactions with purified recombinant proteins. Each binding partner of the bi-molecular interaction is coupled either to donor or to acceptor beads. The technology is based on the quantifiable transfer of oxygen singlets from donor to acceptor microbeads brought together by a specific interaction between the partners. We identified the conserved interaction between WW domains of cellular ubiquitin ligases of the Nedd4 family and a short peptide motif (PPxY) present in several structural and non-structural viral proteins as a potential drug target. Using an AlphaScreen assay recapitulating the interaction between Nedd4.2 and the PPxY motif of the adenoviral capsid protein VI, we screened a library of small molecules and identified specific inhibitors of this interaction.
Intracellular Transport of Viral Structures, MFP CNRS UMR 5234, University of Bordeaux, 146 rue Leo Saignat, Bat. 3A, 3eme etage, Bordeaux, 33076, France.
Sisley
S
Austin
BMY Screen, 229 Cours De L'argonne, Bordeaux, 33076, France.
Saïd
S
Taouji
Inserm ERL440 "Oncogenesis Stress Signaling", Université Rennes 1, Rennes, France.|Centre de Lutte Contre le Cancer Eugène Marquis, Rennes, France.
Eric
E
Chevet
Intracellular Transport of Viral Structures, MFP CNRS UMR 5234, University of Bordeaux, 146 rue Leo Saignat, Bat. 3A, 3eme etage, Bordeaux, 33076, France. harald.wodrich@u-bordeaux.fr.
Harald
H
Wodrich
Intracellular Transport of Viral Structures, MFP CNRS UMR 5234, University of Bordeaux, 146 rue Leo Saignat, Bat. 3A, 3eme etage, Bordeaux, 33076, France.
Fabienne
F
Rayne
D000256:Adenoviridae / Q000235:genetics; D054884:Host-Pathogen Interactions / Q000235:genetics / Q000502:physiology*; D000075702:Nedd4 Ubiquitin Protein Ligases / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D026082:Singlet Oxygen / Q000378:metabolism
D011994:Recombinant Proteins; D026082:Singlet Oxygen; D000075702:Nedd4 Ubiquitin Protein Ligases
384-Well plate; Adenovirus; AlphaScreen®; Antiviral; High-throughput; Protein-Protein interactions; Small-molecule inhibitors; Ubiquitin ligase Nedd4; [LP]PxY motif