Department of Chemistry and Center for Drug Discovery , Virginia Tech , Blacksburg , Virginia 24060 , United States.
Basic Research Laboratory , National Cancer Institute , Frederick , Maryland 21702 , United States.
J Med Chem. 2018 Nov 8;61(21):9611-9620. doi: 10.1021/acs.jmedchem.8b01076. Epub 2018 Oct 18.
We synthesized and screened a unique 46 656-member library composed of unnatural amino acids that revealed several hits against RRE IIB RNA. Among the hit peptides identified, peptide 4A5 was found to be selective against competitor RNAs and inhibited HIV-1 Rev-RRE RNA interaction in cell culture in a p24 ELISA assay. Biophysical characterization in a ribonuclease protection assay suggested that 4A5 bound to the stem-loop region in RRE IIB while SHAPE MaP probing with 234 nt RRE RNA indicated additional interaction with secondary Rev binding sites. Taken together, our investigation suggests that HIV replication is inhibited by 4A5 blocking binding of Rev and subsequent multimerization.
我们合成并筛选了一个独特的 46656 成员的非天然氨基酸文库，该文库揭示了几种针对 RRE IIB RNA 的有效物质。在所鉴定的有效物质肽中，肽 4A5 被发现对竞争 RNA 具有选择性，并在 p24 ELISA 测定中抑制细胞培养中的 HIV-1 Rev-RRE RNA 相互作用。在核糖核酸酶保护测定中的生物物理特性表明，4A5 与 RRE IIB 的茎环区域结合，而 234nt RRE RNA 的 SHAPE MaP 探测则表明与二级 Rev 结合位点的额外相互作用。总之，我们的研究表明，HIV 复制被 4A5 阻止 Rev 结合和随后的多聚化所抑制。
