title: Functional analysis of backbone cyclic peptides bearing the arm domain of the HIV-1 Rev protein: characterization of the karyophilic properties and inhibition of Rev-induced gene expression.
abstract: This work describes the synthesis and activity of a novel backbone cyclic (BC) peptide library based on the sequence of the HIV-1 Rev arginine-rich motif (ARM). All the peptides in the library possess the same sequence but differ in their ring-moiety properties. The BC peptides were synthesized using simultaneous multiple-peptide synthesis and were fully assembled using bis(trichloromethyl)carbonate as a coupling agent. All the peptides in the library had inhibitory effects on the binding of Rev-GFP to importin beta in vitro. Studies performed with one of the BC Rev-ARM analogues, Rev-13, demonstrated that, like its parental linear peptide, it is karyophilic; i.e., it is able to mediate the nuclear import of conjugated bovine serum albumin (BSA) molecules. The cell penetrating properties of the BC peptides were assessed utilizing an ELISA-based system. This assay provides a quantitative evaluation of cell penetration. Most of the peptides from the library were able to penetrate intact Colo-205 cells to varying degrees. Furthermore, these BC peptides were able to carry BSA into intact Colo-205 cells. In addition to its cell penetrating and binding properties, the BC Rev-13 analogue inhibited Rev-induced gene expression in HeLa cells by 60-70% in the low micromolar range and exhibited no cell toxicity. The potential of BC peptides bearing ARM domains as lead compounds for the production of anti-HIV drugs is discussed.
authors.affiliation: Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 9104, Israel.
authors.forename: Elana
authors.initials: E
authors.lastname: Hariton-Gazal
authors.forename: Joseph
authors.initials: J
authors.lastname: Rosenbluh
authors.forename: Nechama
authors.initials: N
authors.lastname: Zakai
authors.forename: Gil
authors.initials: G
authors.lastname: Fridkin
authors.forename: Ruth
authors.initials: R
authors.lastname: Brack-Werner
authors.forename: Horst
authors.initials: H
authors.lastname: Wolff
authors.forename: Christian
authors.initials: C
authors.lastname: Devaux
authors.forename: Chaim
authors.initials: C
authors.lastname: Gilon
authors.forename: Abraham
authors.initials: A
authors.lastname: Loyter
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine; D001665:Binding Sites; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D015694:Gene Products, rev / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D019151:Peptide Library; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000494:pharmacology; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D054321:rev Gene Products, Human Immunodeficiency Virus
chemical_list: D015694:Gene Products, rev; D019151:Peptide Library; D010456:Peptides, Cyclic; D011994:Recombinant Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus; D006160:Guanosine Triphosphate; D001120:Arginine