title: Correlation between shiftide activity and HIV-1 integrase inhibition by a peptide selected from a combinatorial library.
abstract: The human immunodeficiency virus type 1 (HIV-1) integrase (IN) protein is an emerging target for the development of anti-HIV drugs. We recently described a new approach for inhibiting IN by "shiftides"--peptides that inhibit the protein by shifting its oligomerization equilibrium from the active dimer to the inactive tetramer. In this study, we used the yeast two-hybrid system with the HIV-1 IN as a bait and a combinatorial peptide aptamer library as a prey to select peptides of 20 amino acids that specifically bind IN. Five non-homologous peptides, designated as IN-1 to IN-5, were selected. ELISA studies confirmed that IN binds the free peptides. All the five peptides interact with IN with comparable affinity (K(d approximately )10 microM), as was revealed by fluorescence anisotropy studies. Only one peptide, IN-1, inhibited the enzymatic activity of IN in vitro and the HIV-1 replication in cultured cells. In correlation, fluorescence anisotropy binding experiments revealed that of the five peptides, only the inhibitory IN-1 inhibited the DNA binding of IN. Analytical gel filtration experiments revealed that only the IN-1 and not the four other peptides shifted the oligomerization equilibrium of IN towards the tetramer. Thus, the results show a distinct correlation between the ability of the selected peptides to inhibit IN activity and that to shift its oligomerization equilibrium.
authors.affiliation: Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
authors.forename: Ayelet
authors.initials: A
authors.lastname: Armon-Omer
authors.forename: Aviad
authors.initials: A
authors.lastname: Levin
authors.forename: Zvi
authors.initials: Z
authors.lastname: Hayouka
authors.forename: Karin
authors.initials: K
authors.lastname: Butz
authors.forename: Felix
authors.initials: F
authors.lastname: Hoppe-Seyler
authors.forename: Shoshana
authors.initials: S
authors.lastname: Loya
authors.forename: Amnon
authors.initials: A
authors.lastname: Hizi
authors.forename: Assaf
authors.initials: A
authors.lastname: Friedler
authors.forename: Abraham
authors.initials: A
authors.lastname: Loyter
mesh_terms: D000595:Amino Acid Sequence; D052158:Aptamers, Peptide / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D020650:Combinatorial Chemistry Techniques; D019427:HIV Integrase / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D019428:HIV Integrase Inhibitors / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008214:Lymphocytes / Q000187:drug effects / Q000821:virology; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D011485:Protein Binding / Q000187:drug effects; D020836:Protein Structure, Quaternary; D013997:Time Factors; D020798:Two-Hybrid System Techniques; D014779:Virus Replication / Q000187:drug effects
chemical_list: D052158:Aptamers, Peptide; D019428:HIV Integrase Inhibitors; D010455:Peptides; D019427:HIV Integrase