title: Screening for antiviral inhibitors of the HIV integrase-LEDGF/p75 interaction using the AlphaScreen luminescent proximity assay.
abstract: Small-molecule inhibitors of HIV integrase (HIV IN) have emerged as a promising new class of antivirals for the treatment of HIV/AIDS. The compounds currently approved or in clinical development specifically target HIV DNA integration and were identified using strand-transfer assays targeting the HIV IN/viral DNA complex. The authors have developed a second biochemical assay for identification of HIV integrase inhibitors, targeting the interaction between HIV IN and the cellular cofactor LEDGF/p75. They developed a luminescent proximity assay (AlphaScreen) designed to measure the association of the 80-amino-acid integrase binding domain of LEDGF/p75 with the 163-amino-acid catalytic core domain of HIV IN. This assay proved to be quite robust (with a Z' factor of 0.84 in screening libraries arrayed as orthogonal mixtures) and successfully identified several compounds specific for this protein-protein interaction.
authors.affiliation: Department of Virology, Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Hou
authors.forename: Debra E
authors.initials: DE
authors.lastname: McGuinness
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Prongay
authors.forename: Boris
authors.initials: B
authors.lastname: Feld
authors.forename: Paul
authors.initials: P
authors.lastname: Ingravallo
authors.forename: Robert A
authors.initials: RA
authors.lastname: Ogert
authors.forename: Charles A
authors.initials: CA
authors.lastname: Lunn
authors.forename: John A
authors.initials: JA
authors.lastname: Howe
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D004353:Drug Evaluation, Preclinical; D019427:HIV Integrase / Q000187:drug effects* / Q000378:metabolism; D019428:HIV Integrase Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D019428:HIV Integrase Inhibitors; D036341:Intercellular Signaling Peptides and Proteins; C119380:lens epithelium-derived growth factor; D019427:HIV Integrase